GAPDH Recombinant Protein Antigen

• Reactivity: Hu
• Applications: AC

GAPDH Recombinant Protein Antigen Summary

• Description: A recombinant protein antigen with a N-terminal His6-ABP tag corresponding to human GAPDH.

  Source: E. coli

  Amino Acid Sequence: EKPAKYDDIKKVVKQASEGPLKGILGYTEHQVVSSDFNSDTHSSTFDAG

  Fusion Tag: N-terminal His6ABP (ABP = Albumin Binding Protein derived from Streptococcal Protein G)

  This product is intended to be used as a blocking antigen for antibody competition assays. Any other use of this antigen is done at the risk of the user. The use of this product for commercial production is strictly prohibited. Please contact technical support if you have any questions.

• Source: E. coli
• Protein/Peptide Type: Recombinant Protein Antigen
• Gene: GAPDH
• Purity: >80% by SDS-PAGE and Coomassie blue staining

Applications/Dilutions

• Dilutions:
  • Antibody Competition 10 - 100 molar excess
• Application Notes: This recombinant antigen is only intended to be used as a blocking agent to confirm antibody specificity with the corresponding antibody, catalog number NBP2-56153.

  It is purified by IMAC chromatography, and the expected concentration is greater than 0.5 mg/ml.

  For current lot information, including availability, please contact our technical support team click nb-technical@bio-techne.com

• Theoretical MW: 23 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.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS and 1M Urea, pH 7.4.
• Preservative: No Preservative
• Purity: >80% by SDS-PAGE and Coomassie blue staining

Alternate Names for GAPDH Recombinant Protein Antigen

• 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. Peptides and proteins are guaranteed for 3 months from date of receipt.

FAQs for GAPDH Recombinant Protein Antigen (NBP2-56153PEP). (Showing 1 - 7 of 7 FAQ).

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