GAPDH Antibody (NB615)

Simple Western: GAPDH Antibody (NB615) [NB300-285] - 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. Note: band observed higher than predicted 36 kDa molecular weight.

Immunocytochemistry/Immunofluorescence: GAPDH Antibody (NB615) [NB300-285] - Confocal immunofluorescence of HeLa cells using GAPDH antibody at 1:10 (green). Nuclei were counterstained with DAPI (blue).

Western Blot: GAPDH Antibody (NB615) [NB300-285] - Analysis of 40ug HeLa whole cell lysates using NB300-285 at a dilution of 1:100. Note: band observed higher than predicted 36 kDa molecular weight.

Western Blot: GAPDH Antibody (NB615) [NB300-285] - Loss of Ramp2 alters mRNA expression of RAMP2-associating GPCRs, including Pthr1. PTH1R protein expression by immunoblot of e13.5 tissue lysates from wild type and Ramp2-/- placentas. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0181597), licensed under a CC-BY license.

Western Blot: GAPDH Antibody (NB615) [NB300-285] - Analysis of HeLa, Jurkat, and COS lysate using GAPDH (NB300-285) antibody at 1:100. Theoretical molecular weight: 36 kDa.

Product Discontinued

GAPDH Antibody (NB615) Summary

• Immunogen: GAPDH antibody raised against full length human GAPDH from erythrocytes. [UniProt# P04406]
• Localization: Cytoplasmic
• Marker: Cytosolic Marker
• Predicted Species: Rat (94%), Porcine (96%), Bovine (94%), Rabbit (95%), Chicken (94%). Backed by our 100% Guarantee.
• Isotype: IgM
• Clonality: Monoclonal
• Host: Mouse
• Gene: GAPDH
• Purity: IgM purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:100-1:500
  • Simple Western 1:50
  • Western Blot 1:1000-1:5000
• Application Notes: This GAPDH antibody is useful for Western blot and Immunocytochemistry/Immunofluorescence. In Western blot a band is observed at approx. 36 kDa (predicted molecular weight: 40.2 kDa). In ICC/IF, cytoplasmic staining was observed in HeLa cells.
  
  See Simple Western Antibody Database for Simple Western validation: Separated by Size-All, antibody dilution of 1:50. Apparent MW in kDa on Simple Western was 39kDa; matrix was 12-230 kDa.
• 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.
• Control:

  GAPDH Overexpression Lysate

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.07% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: IgM purified

Alternate Names for GAPDH Antibody (NB615)

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

Background

Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) is a ubiquitous enzyme involved in glycolysis, converting glyceraldehyde-3-phosphate into 1,3 diphosphoglycerate. This constitutively expressed, homotetramer protein can be found in the nucleus and cytoplasm, and the monomer has a theoretical molecular weight of 36 kDa. Known as a housekeeping gene, GAPDH routinely serves as a control for real-time PCR (RT-PCR) and as a loading control for Western Blots (1-3). In addition to its role in glycolysis, GAPDH has multiple cellular functions including membrane trafficking, transcription activation, apoptosis, DNA repair, and has been implicated in various pathologies such as cancer and neurodegenerative diseases including Alzheimer's and Huntington's (4,5).

References

1) Barber RD, Harmer DW, Coleman RA, Clark BJ. (2005) GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics. 21(3):389-95. PMID: 15769908

2) Jia Y, Takimoto K. (2006) Mitogen-activated protein kinases control cardiac KChIP2 gene expression. Circ Res. 98(3):386-93. PMID: 16385079

3) Godsel LM, Hsieh SN, Amargo EV, Bass AE, Pascoe-McGillicuddy LT, Huen AC, Thorne ME, Gaudry CA, Park JK, Myung K, Goldman RD, Chew TL, Green KJ. (2005) Desmoplakin assembly dynamics in four dimensions: multiple phases differentially regulated by intermediate filaments and actin. J Cell Biol. 171(6):1045-59. PMID: 16365169

4) Sirover MA1. (1999) New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. Biochim Biophys Acta. 1432(2): 159-84. PMID: 10407139

5) Tristan C, Shahani N, Sedlak TW, Sawa A. (2011) The diverse functions of GAPDH: views from different subcellular compartments. Cell Signal. 23(2):317-23. PMID: 20727968

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 (NB300-285)(11)

Publications using NB300-285

• Kechele DO, Blue RE, Zwarycz B et al. Orphan Gpr182 suppresses ERK-mediated intestinal proliferation during regeneration and adenoma formation J. Clin. Invest 2017-01-17 [PMID: 28094771] (WB, Mouse)
• Shirato K, Ujike M, Kawase M, Matsuyama S. Identification of CCL2, RARRES2 and EFNB2 as host cell factors that influence the multistep replication of respiratory syncytial virus. Virus Res. 2015-08-12 [PMID: 26277777] (WB, Human)
• Gu Y, Bouwman P, Greco D et al. Suppression of BRCA1 sensitizes cells to proteasome inhibitors. Cell Death Dis 2014-12-18 [PMID: 25522274] (WB)
• Rajala HL, Eldfors S, Kuusanmaki H et al. Discovery of somatic STAT5b mutations in large granular lymphocytic leukemia. Blood 2013-04-17 [PMID: 23596048]
• Ichiki T, Boerrigter G, Huntley BK et al. Differential Expression of the Pro-Natriuretic Peptide Convertases Corin and Furin in Experimental Heart Failure and Atrial Fibrosis Am J Physiol Regul Integr Comp Physiol 2012-11-14 [PMID: 23152112] (WB, Canine, Human)
• Chiappinelli KB, Haynes BC, Brent MR et al. Reduced DICER1 elicits an interferon response in endometrial cancer cells. Mol Cancer Res. 2012-03-01 [PMID: 22252463] (WB, Human)
• George, J et al. Restoration of tissue factor pathway inhibitor-2 in a human glioblastoma cell line triggers caspase-mediated pathway and apoptosis. Clin Cancer Res;13(12):3507-17. 2007-06-15 [PMID: 17575213] (WB, Human)
• Sankowski R. Effects of the MAP kinase inhibitor CNI-1493 on Amyloid beta-treated primary and mikrogliare neuronal cells Thesis, Department of Medicine, Philipps-University Marburg. 2012-11-01 (WB, Mouse)
• Sengupta S, Wang H, Yang C et al. Ligand-induced gene activation is associated with oxidative genome damage whose repair is required for transcription Proc. Natl. Acad. Sci. U.S.A. 2020-08-21 [PMID: 32826329]
• Ramsingh G, Koboldt DC, Trissal M et al. Complete characterization of the microRNAome in a patient with acute myeloid leukemia. Blood. 2010-12-09 [PMID: 20876853] (WB, Human)
• Shen, L et al. Drug sensitivity prediction by CpG island methylation profile in the NCI-60 cancer cell line panel. Cancer Res;67(23):11335-43. 2007-12-01 [PMID: 18056460]

Reviews for GAPDH Antibody (NB300-285) (3)

Average Rating: 4.7

(Based on 3 reviews)

We have 3 reviews tested in 1 species: Human.

reviewed by: Bryan Tinsley ICC Human 06/10/2014

Summary

• Application: Immunocytochemistry
• Species: Human

reviewed by: Bryan Tinsley WB Human 06/10/2014

Summary

• Application: Western Blot
• Species: Human

reviewed by: Bryan Tinsley ICC Human 05/22/2014

Summary

• Application: Immunocytochemistry
• Species: Human

FAQs for GAPDH Antibody (NB300-285). (Showing 1 - 7 of 7 FAQs).

1. We need a loading control antibody for WB working with HeLa cell line, will this GAPDH antibody work for that?
   • 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 interested in. 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.
7. I wanted to know which of the two housekeeping genes B-actin or GAPDH can be used for best results. I intend to do an experiment to determine expression of IL-6 and TNF-alpha in liver and kidney tissues.
   • For homogenized tissue, beta-actin and GAPDH are both fine.

Bioinformatics

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