WB (Western Blot)
(Western Blot analysis of GAPDH expression in transfected 293T cell line by GAPDH monoclonal antibody (M01), clone 3C2.Lane 1: GAPDH transfected lysate (36.1 KDa).Lane 2: Non-transfected lysate.)
Mouse GAPDH Monoclonal Antibody | anti-GAPDH antibody
GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase, G3PD, GAPD, MGC88685) (PE)
WB (Western Blot)
(Western Blot analysis of GAPDH expression in transfected 293T cell line by GAPDH monoclonal antibody (M01), clone 3C2.Lane 1: GAPDH transfected lysate (36.1 KDa).Lane 2: Non-transfected lysate.)
WB (Western Blot)
(Western Blot analysis of GAPDH expression in transfected 293T cell line by GAPDH monoclonal antibody (M01), clone 3C2.Lane 1: GAPDH transfected lysate (36.1 KDa).Lane 2: Non-transfected lysate.)
WB (Western Blot)
(GAPDH monoclonal antibody (M01), clone 3C2. Western Blot analysis of GAPDH expression in Raw 264.7.)
WB (Western Blot)
(GAPDH monoclonal antibody (M01), clone 3C2. Western Blot analysis of GAPDH expression in Raw 264.7.)
WB (Western Blot)
(GAPDH monoclonal antibody (M01), clone 3C2. Western Blot analysis of GAPDH expression in PC-12.)
WB (Western Blot)
(GAPDH monoclonal antibody (M01), clone 3C2. Western Blot analysis of GAPDH expression in PC-12.)
WB (Western Blot)
(GAPDH monoclonal antibody (M01), clone 3C2. Western Blot analysis of GAPDH expression in NIH/3T3.)
WB (Western Blot)
(GAPDH monoclonal antibody (M01), clone 3C2. Western Blot analysis of GAPDH expression in NIH/3T3.)
WB (Western Blot)
(GAPDH monoclonal antibody (M01), clone 3C2. Western Blot analysis of GAPDH expression in HeLa (Cat # L013V1).)
WB (Western Blot)
(GAPDH monoclonal antibody (M01), clone 3C2. Western Blot analysis of GAPDH expression in HeLa (Cat # L013V1).)
WB (Western Blot)
(GAPDH monoclonal antibody (M01), clone 3C2 Western Blot analysis of GAPDH expression in A-431 (Cat # L015V1).)
WB (Western Blot)
(GAPDH monoclonal antibody (M01), clone 3C2 Western Blot analysis of GAPDH expression in A-431 (Cat # L015V1).)
NCBI and Uniprot Product Information
Observed: 36 kDa
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Product Notes
The GAPDH gapdh (Catalog #AAA26650) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's GAPDH can be used in a range of immunoassay formats including, but not limited to, WB (Western Blot). Applications are based on unconjugated antibody. Researchers should empirically determine the suitability of the GAPDH gapdh 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 "GAPDH, Monoclonal 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.Frequently Asked Questions
Is GAPDH antibody commonly used as a loading control in Western blot?
Yes, GAPDH (Glyceraldehyde-3-phosphate Dehydrogenase) is one of the three most widely used housekeeping protein loading controls in Western blotting. Its constitutive high expression across tissues and relative stability under most conditions make GAPDH antibodies standard tools for normalizing protein loading and quantifying protein band intensities.
Can a mouse GAPDH antibody be used for immunofluorescence or IHC?
Yes, GAPDH antibodies are suitable for immunofluorescence (IF) and immunohistochemistry (IHC) applications, enabling visualization of GAPDH distribution within cells and tissues. However, GAPDH's ubiquitous cytoplasmic localization typically results in diffuse staining patterns, limiting its utility compared to Western blot applications.
Does GAPDH expression remain stable across different tissues and conditions?
GAPDH expression is generally stable across most tissues and conditions, making it suitable as a loading control. However, recent studies reveal GAPDH regulation varies under certain conditions (ischemia, metabolic stress, specific diseases), requiring validation for use in novel experimental contexts rather than assuming universal stability.
What’s the difference between GAPDH and other housekeeping protein antibodies?
GAPDH (~37 kDa), beta-actin (~42 kDa), and beta-tubulin (~55 kDa) differ in molecular weight and tissue distribution patterns. GAPDH shows superior stability in some neuroscience contexts; beta-tubulin performs better in adipose tissue; beta-actin offers a broader working range. Choice depends on specific experimental requirements and sample types.
Can GAPDH be used to normalize gene or protein expression data?
Yes, GAPDH serves as a reference gene/protein for normalizing qPCR and Western blot data, respectively. GAPDH-normalized gene expression ratios enable comparison of target gene expression across different samples, conditions, and tissues, accounting for variations in starting RNA/protein quantities and extraction efficiency.
Is this GAPDH antibody suitable for quantitative protein assays?
Yes, GAPDH antibodies enable quantitative protein assays via Western blotting, ELISA, and immunofluorescence. GAPDH's abundant expression provides strong signals facilitating accurate quantification across a wide linear range, though researchers should validate GAPDH stability in their specific experimental model before quantitative applications.
Is mouse GAPDH antibody suitable as a loading control for Western blot?
Yes, mouse GAPDH is a standard, well-validated loading control for Western blots. GAPDH (36 kDa) catalyzes glycolytic oxidative phosphorylation and is universally expressed across tissues. Its expression remains relatively stable across most conditions, making it ideal for normalizing protein loading variability. Standard dilutions (1:10,000) yield strong, consistent bands. AAA Biotech's mouse GAPDH antibody is optimized for mammalian lysates; verify species reactivity for mouse-derived samples.
Can this GAPDH antibody be used for immunofluorescence or IHC?
Yes, GAPDH antibodies are suitable for immunofluorescence (IF) and immunohistochemistry (IHC). GAPDH is ubiquitously expressed in all mouse tissues, producing a robust signal. In IF, GAPDH shows cytoplasmic staining (primary cellular localization) with an optional nuclear signal under oxidative stress. For IHC on mouse tissues, standard protocols work well. However, GAPDH can translocate to the nucleus under stress conditions, potentially complicating interpretation if stress response is studied.
Does GAPDH expression stay stable across different tissues and treatments?
Generally stable, but with caveats. GAPDH expression is relatively constant across normal tissues (liver, muscle, brain, kidney). However, some pathological conditions alter GAPDH levels: Alzheimer's disease shows decreased brain GAPDH; alcoholic hepatitis increases liver GAPDH; adipose tissue shows variability. Additionally, GAPDH translocates to the nucleus under oxidative stress. For stressed samples, validate GAPDH stability experimentally or use alternative housekeeping proteins (18S, tubulin, actin variants).
How does GAPDH compare to other housekeeping protein markers?
GAPDH ranks among the best housekeeping proteins alongside β-actin and β-tubulin. GAPDH advantages: multiple cellular functions (glycolysis, apoptosis, transcription), ubiquitous expression, stable across tissues. However, GAPDH shows variability in specific conditions (Alzheimer's, neuronal injury, adipose tissue). β-actin and β-tubulin show similar condition-dependent variability. Optimal practice: use multiple housekeeping proteins (GAPDH + actin) or employ stain-free gel technology for quantitation without protein-dependent normalization.
What dilution is recommended for best results in flow cytometry or WB?
For Western blot: typically 1:10,000 primary antibody dilution in blocking solution (1:500-1:1000 also used). Titrate the antibody to find the optimal dilution balancing signal intensity against the background. For flow cytometry: GAPDH is not commonly used as GAPDH is primarily cytoplasmic; surface markers (CD45, etc.) are preferred. If intracellular GAPDH flow cytometry is needed, use 5-10 μL conjugated antibody per 10⁶ cells after fixation/permeabilization.
Is this antibody validated for quantitative protein assays or normalization?
Yes, GAPDH antibodies are validated for quantitative protein normalization in Western blots. GAPDH levels remain relatively constant, enabling reliable background correction. Signal intensity scales linearly with protein loading. However, for absolute quantification in pathological states (Alzheimer's, obesity, inflammation), validate GAPDH stability first or employ ratiometric approaches using multiple housekeeping proteins. Stain-free gels offer superior quantitation without protein-dependent normalization.
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