Western Blot: GAPDH Antibody [NB300-324] - Detection of Human and Mouse GAPDH by Western Blot. Samples: Whole cell lysate (50 ug) from HeLa, 293T, Jurkat, mouse TCMK-1, and mouse NIH3T3 cells prepared using NETN lysis ...read more
Immunocytochemistry/ Immunofluorescence: GAPDH Antibody [NB300-324] - GAPDH antibody was tested in Hela cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).
Immunohistochemistry-Paraffin: GAPDH Antibody [NB300-324] - IHC detection of GAPDH in formalin-fixed paraffin-embedded mouse squamous cell carcinoma tissue sections with NB300-324 at a dilution of 1:200.
Western Blot: GAPDH Antibody [NB300-324] - Detection of Human and Mouse GAPDH Whole cell lysate from HeLa (5, 15 and 50 mcg) and mouse NIH3T3 (M; 50 mcg). Affinity purified rabbit anti-GAPDH antibody used at 0.04 ...read more
Immunohistochemistry-Paraffin: GAPDH Antibody [NB300-324] - IHC detection of GAPDH in formalin-fixed paraffin-embedded human lung carcinoma tissue sections with NB300-324 at a dilution of 1:200
Proximity Ligation Assay: GAPDH Antibody [NB300-324] - Image showing representative results for Proximity Ligation Assay (PLA ) using three color fluorescence, including DAPI stained nuclei (blue), phalloidin stained ...read more
Simple Western: GAPDH Antibody [NB300-324] - Simple Western lane view shows a specific band for GAPDH in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation ...read more
This GAPDH antibody is useful for Immunohistochemistry-paraffin, Immunocytochemistry/Immunofluorescence and Western blot applications. In WB, a specific band is observed at approx. 36 kDa molecular weight position. In Immunohistochemistry, epitope retrieval with citrate buffer pH6.0 is recommended for formalin-fixed paraffin-embedded tissue sections.
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
36 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Human and mouse. Based upon 100% sequence identity, this antibody is predicted to react with Rat, Chicken, Turkey, Guinea pig, Pig, Rhesus Monkey, Gorilla, Chimpanzee, West Indian ocean coelacanth, Neobatrachia, Sauropsida, Common iguana, Black-banded sea krait, Domestic pigeon and Indian bull frog.
Packaging, Storage & Formulations
Store at 4C. Do not freeze.
TBS and 0.1% BSA
0.09% Sodium Azide
Immunogen affinity purified
Alternate Names for GAPDH Antibody
aging-associated gene 9 protein
GAPDPeptidyl-cysteine S-nitrosylase GAPDH
glyceraldehyde 3-phosphate dehydrogenase
GAPDH (glyceraldehyde-3-phosphate dehydrogenase or GAPD) is a key enzyme in glycolytic pathway, wherein it catalyzes the first step by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. It localizes mainly in the cytoplasm from where it translocates to nucleus following S-nitrosylation and interaction with SIAH1. Nuclear GAPDH implicates in transcription, RNA transport, DNA replication and apoptosis, via its nitrosylase activity which mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2, PRKDC etc. GAPDH also regulates the organization/assembly of cytoskeleton and facilitates CHP1-dependent microtubule - membrane associations. It is a component of GAIT (gamma interferon-activated inhibitor of translation) complex which mediates IFN-gamma-induced transcript-selective translation inhibition in inflammation. Because of its expression as housekeeping protein in most cell types, GAPDH is often used as a control molecule in various genes expression studies, however, recent evidence has shown the association of its altered expression with neurodegenerative pathologies such as Huntington disease, Alzheimer's disease etc., and elevated GAPDH mRNA/protein expression levels have been seen in pancreatic, lung and prostate cancers.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
FAQs for GAPDH Antibody (NB300-324). (Showing 1 - 6 of 6 FAQs).
We need a loading control antibody for WB working with HeLa cell line, will this GAPDH antibody work for that?
Well GAPDH is expressed in HeLa cell lines but whether or not it is the best option for a loading control antibody depends on the size of the protein you are lookin at. GAPDH antibody is a good choice for low to mid MW targets as GAPDH detects around 35 kDa.
I want to know why GAPDH is used as a loading control antibody in WB techniques.
Our GAPDH antibody makes a good loading control because GAPDH is a housekeeping gene essential to metabolism that is globally expressed in most tissue types and cell lines.
I need a suitable loading control antibody that I can use in western blot of rat nerve extracts, which do you recommend?
This GAPDH antibody (NB300-221) is a good choice for a loading control antibody and has been cited in over 180 publications and used on rat lysates with good results.
I'm looking for a loading control between 24kDa - 65kDa for use in simple western, we tried an actin antibody we already had but it didn't work. What can you recommend?
Our GAPDH antibody NB300-221 has been validated in simple western and detects around 35 kDa so I think it would be a good choice for a simple western loading control antibody based on your size requirements.
What secondary antibody should I use for visualization of the GAPDH antibody?
This GAPDH antibody is raised in mouse so you would want to use an anti-mouse secondary with the dye of your choice.
Do you offer a smaller size for the GAPDH antibodies?
We offer sample sizes for many of our GAPDH loading control antibodies including NB300-221.
GAPDH: More than a housekeeping gene GAPDH is a 146kD tetramer glycolytic pathway metabolic enzyme composed of four 30-40 kDa subunits. It is responsible for reversibly phosphorylating its substrate glyceraldehyde 3-phosphate within the glycolytic pathway. Apart from its role in gl... Read full blog post.
The GAPDH Antibody in Western blot Assays The loading controls on our antibody database are widely used in gel electrophoresis and Western blotting studies. Products like the GAPDH antibody detect "housekeeping" proteins which are abundantly distributed in cells. This makes them useful for ch... Read full blog post.
New Primers Available for ChIP Procedures Novus has recently released six new primer sets designed for use in real-time PCR DNA amplification of housekeeping, silent, or heterochromatin associated proteins. Specifically, the primers are for GAPDH, RPL30, MyoD1, AFM, alpha Satellite, and Satel... Read full blog post.
The GAPDH Antibody: A ChIP Off the Old Block GAPDH antibodies are commonly used as a loading control, due to GAPDH's widespread function as a house-keeping protein. As a leading scientific research antibody supplier, Novus Biologicals offer a wide range of GAPDH antibody reagents and support pro... Read full blog post.
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