Phospho-specific antibodies’ typical purpose is to enable researchers to detect changes in proteins. They will exclusively bind to the amino acid sequence on a protein that has been phosphorylated (which is both a physical & chemical change) and do not bind to the same amino acid sequence on said protein if it lacks said phosphorylation. This aids in being able to clearly see and understand the data produced from this particular protein modification.
WB (Western Blot) (Western blot analysis of Phospho-p62 Dok (Tyr398) expression in various lysates)
WB (Western Blot) (Western blot analysis of p62 Dok phosphorylation expression in K562 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321767 staining Hela by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IHC (Immunohistochemistry) (AAA321772 at 1/100 staining Human lung tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of Phospho-GRF-1 (Tyr1105) expression in various lysates)
WB (Western Blot) (Western blot analysis of GRF-1 phosphorylation expression in EGF treated 293 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321772 staining 293 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IF (Immunofluorescence) (AAA321774 staining K562 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of p70 S6 Kinase beta phosphorylation expression in EGF treated K562 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
WB (Western Blot) (Western blot analysis of Phospho-p70 S6 Kinase beta (Ser423) expression in various lysates)
WB (Western Blot) (Western blot analysis of Phospho-DARPP-32 (Thr75) expression in various lysates)
IHC (Immunohistochemisry) (AAA321775 at 1/100 staining human Breast cancer tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of DARPP-32 phosphorylation expression in Forskolin treated COS7 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321775 staining Hela by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Western Blot
Purity
From purified rabbit serum by affinity purification via sequential chromatography on phospho-and non-phospho-peptide affinity columns.
Pricing
Application Data (The TFact MYB (Phospho-Ser12) DNA-Binding ELISA detects active MYB (Phospho-Ser12) in Hela and Jurkat Nuclear Extract. The Hela and Jurkat cells were grown 3 days in DMEM and RPMI 1640 with 10% FBS and harvested for nuclear extract.)
ELISA (Enzyme-Linked Immunosorbent Assay (ELISA) for immunogen phosphor-peptide (left) and non-phospho peptide (right), using Anti-Chk2 (Phospho-Thr68) Antibody.)
Application Data (HeLa cells were grown to 90% confluency and were stimulated with PMA 150 ng/mL for 30 minutes. Cells were immediately lysed thereafter and measured for Total Protein Concentration and O.D. 450 nm of Chk2 P-Thr68 versus untreated HeLa lysates.)
Application Data (Jurkat cells were grown to 90% confluency and were stimulated with U.V. 150 J/s and incubated for 1 hr. Cells were immediately lysed thereafter and measured for Total Protein Concentration and O.D. 450 nm of DNA-PK P-Ser2056 versus untreated Jurkat lysates.)
IHC (Immunohiostchemistry) (Immunohistochemical analysis of paraffin-embedded human tonsil. 1, Tris-EDTA,pH9.0 was used for antigen retrieval. 2 Antibody was diluted at 1:200(4 degree overnight.3,Secondary antibody was diluted at 1:200(room temperature, 45min).)
WB (Western Blot) (Western Blot analysis of 1 HeLa cell, 2 LPS 100ng/mL 30min treated ,using primary antibody at 1:1000 dilution. Secondary antibody)
IHC (Immunohiostchemistry) (Immunohistochemical analysis of paraffin-embedded human liver cancer. 1, Antibody was diluted at 1:200(4 degree overnight). 2, Tris-EDTA,pH9.0 was used for antigen retrieval. 3,Secondary antibody was diluted at 1:200(room temperature, 45min).)
WB (Western Blot) (Western Blot analysis of mouse lung , rat lung,using primary antibody at 1:1000 dilution. Secondary antibody)
IHC (Immunohiostchemistry) (Immunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200(4 degree overnight). 2, Tris-EDTA,pH9.0 was used for antigen retrieval. 3,Secondary antibody was diluted at 1:200(room temperature, 45min).)
WB (Western Blot) (Western Blot analysis of 1 A431 treated with LPS, 2 A431,using primary antibody at 1:1000 dilution. Secondary antibody)
Purified from rabbit antiserum by affinity-chromatography using phospho peptide. The antibody against non-phospho peptide was removed by chromatography using corresponding non-phospho peptide.
Purified from rabbit antiserum by affinity-chromatography using phospho peptide. The antibody against non-phospho peptide was removed by chromatography using corresponding non-phospho peptide.
WB (Western Blot) (Western blot analysis of lysates from Jurkat cells treated with starved 24h, using Trk A (Phospho-Tyr680+Tyr681) Antibody.)
ELISA (Enzyme-Linked Immunosorbent Assay (Phospho-ELISA) for Immunogen Phosphopeptide (Phospho-left) and Non-Phosphopeptide (Phospho-right), using Trk A (Phospho-Tyr680+Tyr681) Antibody)
Purified from rabbit antiserum by affinity-chromatography using phospho peptide. The antibody against non-phospho peptide was removed by chromatography using corresponding non-phospho peptide.
Purified from rabbit antiserum by affinity-chromatography using phospho peptide. The antibody against non-phospho peptide was removed by chromatography using corresponding non-phospho peptide.
IF (Immunofluorescence) (Immunofluorescent analysis of Phosphorylation of H2A.X at Serine 139 in A549(upper.untreated or Hydroxyurea-treated) and Hela(lower.untreated or Hydroxyurea-treated) using Phospho-Histone H2A.X (Ser139) mouse mAb (1:400).)
IF (Immunofluorescence) (Immunofluorescent analysis of Phosphorylation of H2A.X at Serine 139 in 3T3 or Hydroxyurea-treated 3T3 cells using Phospho-Histone H2A.X)
WB (Western Blot) (Western blot detection of Phosphorylation of H2A.X at Serine 139 in 3T3 or Hydroxyurea-treated 3T3 cell lysates using Phospho-Histone H2A.X (Ser139) mouse mAb (1:2000 diluted).Predicted band size:15KDa.Observed band size:15KDa.)
WB (Western Blot) (Western blot analysis of extracts from untreated or Hydroxyurea-treated Hela and A549 cells. using Histone H2A.X(Phospho-Ser139) mouse mAb (1:1000 diluted)(upper) or ?-Actin Mouse mAb (200068-8F10)(lower).Predicted band size:15KDa.Observed band size:15KDa.)
IHC (Immunohiostchemistry) (AAA321405 at 1/100 staining Human thyroid cancer tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of extracts of U-2 OS cells, using Phospho-TCTP (Ser46) Antibody.)
WB (Western Blot) (Western blot analysis of extracts of 293T cells, using Phospho-Topoisomerase IIalpha (Ser1469) Antibody.)
IHC (Immunohistochemistry) (AAA321409 at 1/100 staining Mouse spleen tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IHC (Immunohiostchemistry) (AAA321411 at 1/100 staining Human thyroid cancer tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of extracts of NIH/3T3 cells treated with BDNF, using Phospho-TrkA (Tyr490) Antibody.)
IHC (Immunohiostchemistry) (AAA321413 at 1/100 staining Human thyroid cancer tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of extracts of NIH/3T3 cells, using Phospho-Tuberin/TSC2 (Ser939) Antibody.)
IHC (Immunohiostchemistry) (AAA321420 at 1/100 staining Human thyroid cancer tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of extracts of HepG2 cells, using Phospho-Calnexin(Ser564) Antibody.)
IHC (Immunohiostchemistry) (AAA321421 at 1/100 staining Human breast cancer tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of extracts of Jurkat cells, using Phospho-CDK6(Tyr13) Antibody.)
IF (Immunofluorescence) (AAA321427 staining HeLa by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IHC (Immunohistochemistry) (Phospho-Chk1 (Ser345) Antibody for IHC in human brain.)
WB (Western Blot) (Western blot analysis of IKK-beta phosphorylation expression in COS7 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
WB (Western Blot) (Western blot analysis of Phospho-IKK beta (Tyr188) Antibody expression in COS7 cells lysates.The lane on the right is treated with the antigen-specific peptide.)
IHC (Immunohiostchemistry) (AAA321428 at 1/200 staining human liver cancer tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IF (Immunofluorescence) (AAA321428 staining COS7 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of IKK-alpha phosphorylation expression in EGF treated MDA-MB-435 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
WB (Western Blot) (Western blot analysis of Phospho-IKK alpha (Thr23) expression in various lysates)
IF (Immunofluorescence) (AAA321431 staining 293 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IHC (Immunohistochemistry) (AAA321431 at 1/200 staining human colon cancer tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of Dab1 phosphorylation expression in LOVO whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321439 staining HeLa by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of Phospho-Dab1 (Tyr232) expression in various lysates)
Immunocytochemistry, Immunofluorescence, Western Blot
Purity
The Ab is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
WB (Western Blot) (Western blot analysis of Phospho-MSK1 (Ser376) expression in various lysates)
WB (Western Blot) (Western blot analysis of MSK1 phosphorylation expression in HeLa whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
WB (Western Blot) (Western blot analysis of Phospho-EGFR (Thr693) expression in various lysates)
WB (Western Blot) (Western blot analysis of EGFR phosphorylation expression in A431 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IHC (Immunohistochemisry) (AAA321448 at 1/200 staining human colon carcinoma sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IF (Immunofluorescence) (AAA321448 staining SK-OV3 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IF (Immunofluorescence) (AAA321448 staining BT-20 cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37 degree C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37 degree C. A Alexa Fluor 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.)
WB (Western Blot) (Western blot analysis of Phospho-EGFR (Ser1070) expression in various lysates)
WB (Western Blot) (Western blot analysis of EGFR phosphorylation expression in EGF treated SK-OV3 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IHC (Immunohiostchemistry) (AAA321449 at 1/200 staining human lymph node tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IF (Immunofluorescence) (AAA321449 staining SK-OV3 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of Phospho-EGFR (Tyr1172) expression in various lysates)
WB (Western Blot) (Western blot analysis of EGFR phosphorylation expression in EGF treated A431 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321452 staining SK-OV3 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of Estrogen Receptor alpha phosphorylation expression in Estradiol treated MCF7 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IHC (Immunohiostchemistry) (AAA321460 at 1/100 staining human Breast carcinoma tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IF (Immunofluorescence) (AAA321460 staining 293 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of C-RAF phosphorylation expression in K562 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IHC (Immunohistochemisry) (AAA321463 at 1/200 staining human colon carcinoma sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IF (Immunofluorescence) (AAA321463 staining Hela by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of Phospho-C-RAF (Ser259) expression in various lysates)
WB (Western Blot) (Western blot analysis of C-RAF phosphorylation expression in PMA treated NIH-3T3 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
WB (Western Blot) (Western blot analysis of Phospho-C-RAF (Ser338) expression in various lysates)
IF (Immunofluorescence) (AAA321464 staining Hela by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of p53 phosphorylation expression in Adriamycin treated HeLa whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IHC (Immunohistochemisry) (AAA321467 at 1/100 staining Mouse colon tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IF (Immunofluorescence) (AAA321467 staining MDA-MB-435 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IHC (Immunohistochemistry) (IHC analysis of gastric tissue, using Phospho-p53 (Ser366) Antibody.)
WB (Western Blot) (Western blot analysis of HSP27 phosphorylation expression in UV treated HeLa whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IHC (Immunohiostchemistry) (AAA321469 at 1/200 staining human kidney sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IF (Immunofluorescence) (AAA321469 staining HeLa by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of HSP27 phosphorylation expression in Ca2+ treated HeLa whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321470 staining HeLa by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of Amyloid beta A4 phosphorylation expression in HeLa whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321473 staining HeLa by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of Phospho-Amyloid beta A4 (Thr743/668) expression in various lysates)
IHC (Immunohiostchemistry) (AAA321473 at 1/200 staining human kidney tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IF (Immunofluorescence) (AAA321473 staining Hela cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37 degree C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37 degree C. A Alexa Fluor 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.)
WB (Western Blot) (Western blot analysis of Integrin beta3 phosphorylation expression in Insulin treated 293 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321475 staining HepG2 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis of PKCB phosphorylation expression in heatshock treated HeLa whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
WB (Western Blot) (Western blot analysis of Phospho-PKCB (Ser661) Antibody expression in heatshock treated Hela cells lysates.The lane on the right is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321478 staining HeLa by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IHC (Immunohistochemisry) (AAA321479 at 1/100 staining human Brain tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of Phospho-IR (Tyr1355) Antibody expression in Heatshock treated 293 cells lysates.The lane on the right is treated with the antigen-specific peptide.)
WB (Western Blot) (Western blot analysis of IR phosphorylation expression in Heatshock treated 293 whole cell lysates, The lane on the left is treated with the antigen-specific peptide.)
WB (Western Blot) (Western blot analysis on HuvEc cell lysate using Phospho-APC(Ser2054) Antibody. The lane on the left is treated with the antigen-specific peptide.)
IHC (Immunohiostchemistry) (AAA321246 at 1/100 staining human Rectum tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IF (Immunofluorescence) (AAA321246 staining HuvEc by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IF (Immunofluorescence) (AAA321250 staining HuvEc by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
WB (Western Blot) (Western blot analysis on HuvEc cell lysate using Phospho-Nibrin(Ser278) Antibody. The lane on the left is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321250 staining K-562 cells by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) antibody, diluted at 1/600, was used as secondary antibody.)
IF (Immunofluorescence) (AAA321257 staining Hela cells by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) antibody, diluted at 1/600, was used as secondary antibody.)
WB (Western Blot) (Western blot analysis on HeLa cell lysate using Phospho-BTK(Tyr223) Antibody. The lane on the left is treated with the antigen-specific peptide.)
WB (Western Blot) (Western blot analysis on HeLa cell lysate using Phospho-CDC25A(Ser75) Antibody. The lane on the left is treated with the antigen-specific peptide.)
IF (Immunofluorescence) (AAA321275 staining HeLa by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IHC (Immunohistochemistry) (AAA321275 at 1/100 staining human lymph node tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
IF (Immunofluorescence) (AAA321284 staining HepG2 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)
IHC (Immunohiostchemistry) (AAA321284 at 1/100 staining Human breast cancer tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of extracts from JK treated with UV, using Phospho-AKT1/2/3(Tyr315/316/312) Antibody. Lane1 was treated with phospho-blocking peptide, Lane2 was treated with non-phospho-blocking peptide.)
IHC (Immunohiostchemistry) (AAA321287 at 1/100 staining Mouse lung tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of extracts of HeLa serum-starved treated with TPA, using Phospho-FOS (Ser32) Antibody.)
IHC (Immunohiostchemistry) (AAA321294 at 1/100 staining Rat brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
WB (Western Blot) (Western blot analysis of extracts of Ramos serum-starved, using Phospho-BTK(Tyr551) Antibody.)
WB (Western Blot) (Western blot analysis of extracts of NIH/3T3 cells, using Phospho-PKD2(Ser812) Antibody.)
IHC (Immunohistochemistry) (AAA321297 at 1/100 staining Human thyroid cancer tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
From purified rabbit serum by affinity purification via sequential chromatography on phospho-and non-phospho-peptide affinity columns.
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What Are Phospho Antibodies?
Protein phosphorylation is a process where a phosphate group is added to certain amino acid residues of a protein – usually serine (S), threonine (T), or tyrosine (Y) - by enzymes called kinases. This process is integral in controlling cellular signaling, cellular growth, and other biological functions.
Our catalog includes a wide range of phospho-specific antibodies that can accurately detect this important marker. They perform strongly in widely-used laboratory applications such as Western blot, flow cytometry, immunohistochemistry, and immunofluorescence microscopy. We value your trust in us and are committed to providing top-quality products and services. All of our antibodies are guaranteed to work for the applications and species indicated on our website & associated product pages.
What Are The Key Applications of Phospho Antibodies?
1. Western Blotting
One of the first steps a researcher can take in utilizing these phospho-specific antibodies, is to check if the antibody works using a technique referred to as “Western blot”. For those unfamiliar, Western Blot aids in showing whether the protein that the antibody recognizes is appearing at the correct/expected size. These phospho-specific antibodies should also be able to detect changes in the target protein’s phosphorylation (on/off state) when cells are stimulated in certain ways.
2. Staining of Fixed Cells (Immunocytochemistry)
Another routine use of these phospho-specific antibodies, is to test if the antibody is able to demonstrate similar performance when used on fixed cells (intact cells that have been preserved) as it did in the Western blot tests. It is an important aspect in many cases to confirm that the antibody works in actual intact cell samples. Ideally, the method used for cellular fixation should be the same as what is used in pathology labs (like using 10% formalin). To check if the antibody works well in tissue sections (FFPE), researchers will often test it on fixed cells that are processed similar to tissue samples.
3. Specificity Tests Using Peptides
In order to make sure that the antibody is only binding to the right target:
Laboratory technicians will mix the antibody with phospho-peptides (short segments of the protein containing the phosphate group modification).
If the antibody signal disappears, it is confirmation that it is binding to the correct phosphorylated location.
A more robust test is to use both the phosphorylated and non-phosphorylated (dephosphorylated) versions of the protein. The antibody should react only with the phosphorylated one.
Another method sometimes utilized is to treat the sample with an enzyme, such as alkaline phosphatase, that specifically removes phosphate groups. If the antibody signal disappears after this, it also confirms specificity.
4. Genetic Confirmation
As a final step, scientists can genetically manipulate the nucleotide sequence and alter the target protein by removing the exact site where phosphorylation happens. If the antibody no longer appears to detect the modified protein, it is strong evidence supporting the antibody being specific for that phosphorylated site.
Why Buy Phospho Antibodies Through Us?
The production laboratory adheres to strict and consistent protocols prior to releasing any of these phospho-specific antibodies:
Standard methods and proper controls in all tests to ensure high quality.
These antibodies are tested and validated in different cell types and species.
High quality control criterion to ensure each batch is consistent, so you will obtain reliable results every time.
FAQ
1. What Are Phospho-Specific Antibodies?
Phospho-specific antibodies are made to detect proteins only when they have a phosphate group linked to a specific amino acid residue. This empowers scientists understand if a protein is "turned on" or active, based on its phosphorylation state.
2. How to Detect Phosphorylated Proteins in a Western Blot?
To find out if a protein is phosphorylated using Western blot:
Use a phospho-specific antibody that binds only to the phosphorylated form of the protein.
You can also use a “regular” antibody for the same amino acid sequence of the protein that the phospho-specific antibody is binding to (but in this case, this antibody will not bind if there is a phosphate group present) in order to compare how much of it is phosphorylated versus how much is non-phosphorylated (or “total” protein, if the “normal” antibody’s epitopes are non-phospho-site-specific).
3. How to Choose the Best Antibody?
Here are some simple tips to help you pick the right antibody:
Know your target
Match your sample characteristics
Confirm the intended use is appropriate
Check “host” and “type”
Check the “quality” of the presented data/images
Appraise whether the available validation meets your needs
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