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product-image-AAA125544_FCM11.jpg FCM/FACS (Flow Cytometry) (Figure 3. Flow Cytometry analysis of RAW264. 7 cells using anti-TMS1/ASC/PYCARD antibody (AAA125544).Overlay histogram showing RAW264. 7 cells stained with AAA125544 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TMS1/ASC/PYCARD Antibody (AAA125544, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Rabbit anti-Mouse, Rat TMS1/ASC/Pycard Polyclonal Antibody | anti-Pycard antibody

Anti-TMS1/ASC/Pycard Antibody

Gene Names
Pycard; Asc; TNS1; masc; CARD5; TMS-1; 9130417A21Rik
Reactivity
Mouse, Rat
Applications
ELISA, Flow Cytometry, Functional Assay, Immunohistochemistry, Western Blot
Purity
Immunogen affinity purified.
Synonyms
TMS1/ASC/Pycard, Antibody; Anti-TMS1/ASC/Pycard Antibody; Pycard; Asc; Apoptosis-associated speck-like protein containing a CARD; mASC; PYD and CARD domain-containing protein
; PYD and CARD domain containing; anti-Pycard antibody
Ordering
Host
Rabbit
Reactivity
Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Rabbit IgG polyclonal antibody for TMS1/ASC/PYCARD detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Applicable Applications for anti-Pycard antibody
ELISA (Direct ELISA), FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry), WB (Western Blot)
Immunogen
E Coli-derived mouse TMS1/ASC/PYCARD recombinant protein (Position: R3-S193).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

FCM/FACS (Flow Cytometry)

(Figure 3. Flow Cytometry analysis of RAW264. 7 cells using anti-TMS1/ASC/PYCARD antibody (AAA125544).Overlay histogram showing RAW264. 7 cells stained with AAA125544 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TMS1/ASC/PYCARD Antibody (AAA125544, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

product-image-AAA125544_FCM11.jpg FCM/FACS (Flow Cytometry) (Figure 3. Flow Cytometry analysis of RAW264. 7 cells using anti-TMS1/ASC/PYCARD antibody (AAA125544).Overlay histogram showing RAW264. 7 cells stained with AAA125544 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TMS1/ASC/PYCARD Antibody (AAA125544, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IHC (Immunohiostchemistry)

(Figure 2. IHC analysis of TMS1/ASC/PYCARD using anti-TMS1/ASC/PYCARD antibody (AAA125544).TMS1/ASC/PYCARD was detected in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-TMS1/ASC/PYCARD Antibody (AAA125544) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

product-image-AAA125544_IHC13.jpg IHC (Immunohiostchemistry) (Figure 2. IHC analysis of TMS1/ASC/PYCARD using anti-TMS1/ASC/PYCARD antibody (AAA125544).TMS1/ASC/PYCARD was detected in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-TMS1/ASC/PYCARD Antibody (AAA125544) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of TMS1/ASC/PYCARD using anti-TMS1/ASC/PYCARD antibody (AAA125544).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: rat lung tissue lysatesLane 2: rat spleen tissue lysatesLane 3: rat thymus tissue lysatesLane 4: mouse lung tissue lysatesLane 5: mouse spleen tissue lysatesLane 6: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-TMS1/ASC/PYCARD antigen affinity purified polyclonal antibody (Catalog # AAA125544) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for TMS1/ASC/PYCARD at approximately 22KD. The expected band size for TMS1/ASC/PYCARD is at 22KD)

product-image-AAA125544_WB15.jpg WB (Western Blot) (Figure 1. Western blot analysis of TMS1/ASC/PYCARD using anti-TMS1/ASC/PYCARD antibody (AAA125544).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.Lane 1: rat lung tissue lysatesLane 2: rat spleen tissue lysatesLane 3: rat thymus tissue lysatesLane 4: mouse lung tissue lysatesLane 5: mouse spleen tissue lysatesLane 6: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-TMS1/ASC/PYCARD antigen affinity purified polyclonal antibody (Catalog # AAA125544) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for TMS1/ASC/PYCARD at approximately 22KD. The expected band size for TMS1/ASC/PYCARD is at 22KD)
Related Product Information for anti-Pycard antibody
PYCARD, often referred to as ASC (Apoptosis-associated speck-like protein containing a CARD), is a protein that in humans is encoded by the PYCARD gene. This gene encodes an adaptor protein that is composed of two protein-protein interaction domains: a N-terminal PYRIN-PAAD-DAPIN domain (PYD) and a C-terminal caspase-recruitment domain (CARD). The PYD and CARD domains are members of the six-helix bundle death domain-fold superfamily that mediates assembly of large signaling complexes in the inflammatory and apoptotic signaling pathways via the activation of caspase. In normal cells, this protein is localized to the cytoplasm; however, in cells undergoing apoptosis, it forms ball-like aggregates near the nuclear periphery. Two transcript variants encoding different isoforms have been found for this gene.
References
1. Agostini, L., Martinon, F., Burns, K., McDermott, M. F., Hawkins, P. N., Tschopp, J. NALP3 forms an IL-1-beta-processing inflammasome with increased activity in Muckle-Wells autoinflammatory disorder. Immunity 20: 319-325, 2004.
2. Cai, X., Chen, J., Xu, H., Liu, S., Jiang, Q. -X., Halfmann, R., Chen, Z. J. Prion-like polymerization underlies signal transduction in antiviral immune defense and inflammasome activation. Cell 156: 1207-1222, 2014.
3. Conway, K. E., McConnell, B. B., Bowring, C. E., Donald, C. D., Warren, S. T., Vertino, P. M. TMS1, a novel proapoptotic caspase recruitment domain protein, is a target of methylation-induced gene silencing in human breast cancers. Cancer Res. 60: 6236-6242, 2000.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
21,459 Da
NCBI Official Full Name
apoptosis-associated speck-like protein containing a CARD
NCBI Official Synonym Full Names
PYD and CARD domain containing
NCBI Official Symbol
Pycard
NCBI Official Synonym Symbols
Asc; TNS1; masc; CARD5; TMS-1; 9130417A21Rik
NCBI Protein Information
apoptosis-associated speck-like protein containing a CARD
UniProt Protein Name
Apoptosis-associated speck-like protein containing a CARD
UniProt Gene Name
Pycard
UniProt Synonym Gene Names
Asc; mASC
UniProt Entry Name
ASC_MOUSE

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Product Notes

The Pycard pycard (Catalog #AAA125544) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-TMS1/ASC/Pycard Antibody reacts with Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's TMS1/ASC/Pycard can be used in a range of immunoassay formats including, but not limited to, ELISA (Direct ELISA), FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry), WB (Western Blot). Researchers should empirically determine the suitability of the Pycard pycard for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "TMS1/ASC/Pycard, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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