GAPDH Antibody (1A10)

Western Blot: GAPDH Antibody (1A10) [NBP1-47339] - Western blot analysis using anti-GAPDH mAb against Hela (1), A549 (2), A431 (3), MCF-7 (4), K562 (5), Jurkat (6), HL60 (7), SKN-SH (8) and SKBR-3 (9) cell lysate.

Immunocytochemistry/Immunofluorescence: GAPDH Antibody (1A10) [NBP1-47339] - GAPDH was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using 2 ug/mL of mouse anti-GAPDH monoclonal (NB300-328, Novus Biologicals), 2 ug/mL mL of mouse anti- GAPDH monoclonal (NBP1-47339, Novus Biologicals). Cells were stained using the appropriate secondary antibody donkey anti-mouse IgG-NL557 (NL007) and counterstained with DAPI (blue).

Immunohistochemistry-Paraffin: GAPDH Antibody (1A10) [NBP1-47339] - Immunohistochemical analysis of paraffin-embedded human breast carcinoma (left) and kidney carcinoma (right), showing cytoplasmic localization using anti-GAPDH mAb with DAB staining.

Immunocytochemistry/Immunofluorescence: GAPDH Antibody (1A10) [NBP1-47339] - Confocal immunofluorescence analysis of methanol-fixed HepG2 (left) and Hela (right) cells using anti-GAPDH mAb (green), showing cytoplasmic localization. Blue: DRAQ5 fluorescent DNA dye.

Simple Western: GAPDH Antibody (1A10) [NBP1-47339] - 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 system.

• Reactivity: Hu, Mu
• Applications: WB, Simple Western, ELISA, ICC/IF, IHC, IHC-P
• Clone: 1A10
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

GAPDH Antibody (1A10) Summary

• Immunogen: Purified recombinant fragment of human GAPDH expressed in E. coli. [UniProt# P04406]
• Localization: Cytoplasmic
• Marker: Cytosolic Marker
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: GAPDH
• Purity: Ammonium sulfate precipitation

Applications/Dilutions

• Dilutions:
  • Western Blot 1:500-1:2000
  • Simple Western 1:50
  • ELISA 1:10000
  • Immunocytochemistry/Immunofluorescence 1:200-1:1000
  • Immunohistochemistry 1:200-1:1000
  • Immunohistochemistry-Paraffin 1:200-1:1000
• Application Notes: This GAPDH (1A10) antibody is useful for Western blot, Immunohistochemistry on paraffin embedded sections, Immunocytochemistry/Immunofluorescence and ELISA.
  
  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.
• Theoretical MW: 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.
• Positive Control:

  Jurkat Lysate (NB800-PC2)
  A-431 Lysate (NB800-PC12)
  HeLa Lysate (NB800-PC1)
  MCF-7 Lysate (NB820-59465)

Reactivity Notes

Human. Mouse reactivity reported in scientific literature (PMID: 25833141)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.03% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Ammonium sulfate precipitation

Alternate Names for GAPDH Antibody (1A10)

• aging-associated gene 9 protein
• EC 1.2.1
• EC 1.2.1.12
• EC 2.6.99.-
• G3PD
• G3PDH
• GAPDH
• GAPDPeptidyl-cysteine S-nitrosylase GAPDH
• glyceraldehyde 3-phosphate dehydrogenase
• glyceraldehyde-3-phosphate dehydrogenase
• MGC88685

Background

GAPDH is a 146 kDa tetramer composed of four 30-40 kDa subunits. Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) is a metabolic enzyme responsible for catalyzing one step in the glycolytic pathway, the reversible oxidative phosphorylation of glyceraldehyde 3-phosphate. Because GAPDH as a protein expressed in large amounts and which is required at all times for an important house keeping functions, levels of GAPDH mRNA are often used as standards in studies of mRNA expression. Increasingly, scientists are making use of specific antibodies to GAPDH as loading controls for western blotting experiments. Apart from a role in glycolysis, GAPDH may have other roles such as in the activation of transcription. GAPDH is reported to bind to a variety of other proteins, including the amyloid precursor protein, mutations in which cause some forms of Alzheimer's disease, and the polyglutamine tracts of Huntingtin, the protein product aberrant forms of which are causative of Huntington's disease. Associations with actin and tubulin have also be reported. The protein may also have a role in the regulation of apoptosis, and interestingly migrates from the cytoplasm into the nucleus when cells become apoptotic.

Limitations

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.

Publications for GAPDH Antibody (NBP1-47339)(5)

Publications using NBP1-47339

• Hoesel B, Mussbacher M, Dikorman B, Salzmann M. Androgen receptor dampens tissue factor expression via NF-kB and EGR1. J. Thromb. Haemost. 2018 Feb 10 [PMID: 29427323] (Human)
• Li Y, Zhang DW, Lin DQ, Cao LQ. Peroxisome proliferator-activated receptor-gamma inhibits pancreatic cancer cell invasion and metastasis via regulating MMP-2 expression through PTEN. Mol Med Rep. 2015 Aug 12 [PMID: 26299428] (WB)
• Moscovitz O, Ben-Nissan G, Fainer I et al. The Parkinson's-associated protein DJ-1 regulates the 20S proteasome Nat Commun 2015 Apr 02 [PMID: 25833141] (WB, Mouse)
• Zhang Z, Jiang T, Li Q et al. Nodal activates smad and extracellular signal-regulated kinases 1/2 pathways promoting renal cell carcinoma proliferation. Mol Med Rep 2015 Jul [PMID: 25672326]
• Fuzesi-Levi MG, Ben-Nissan G, Bianchi E et al. Dynamic Regulation of the COP9 Signalosome in Response to DNA Damage. Mol. Cell. Biol. 2014 Mar 1 [PMID: 24421388] (WB, Human)

Reviews for GAPDH Antibody (NBP1-47339) (2)

Average Rating: 4.5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Human.

reviewed by: Anonymous WB Human 12/23/2014

Summary

• Application: Western Blot
• Sample Tested: See PMID 24421388
• Species: Human

Blocking

• Blocking Details: See PMID 24421388

Details

• Detection Notes: See PMID 24421388

Comments

• Comments: Independent review from antibodyresource.comPublished in PMID 24421388.

reviewed by: Ranveer Singh WB Human 06/02/2014

Summary

• Application: Western Blot
• Sample Tested: HeLa cells
• Species: Human
• Comments: A good antibody. Can be reused multiple times.

Blocking

• Blocking Details: 5% BSA in TBST for 1 h

Primary Anitbody

• Dilution Ratio: 1:3000, incubated 3h at RT in 5% BSA in TBST

Secondary Antibody

• Secondary Description: Goat anti-mouse IgG HRP (Biorad)
• Secondary Manufacturer Cat#: 170-5047
• Secondary Concentration: 1:10,000

Details

• Detection Notes: ECL reagent was used. Exposure for 10 seconds

Comments

• Comments: A good antibody. Can be reused multiple times.

FAQs for GAPDH Antibody (NBP1-47339). (Showing 1 - 6 of 6 FAQs).

1. 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.
2. 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.
3. 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.
4. 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.
5. 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.
6. 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.

Bioinformatics

• Gene Symbol: GAPDH
• Entrez:
  • 2597(Human)
  • 14433(Mouse)
  • 24383(Rat)
• Uniprot:
  • P04406(Human)