GAPDH Mouse

GAPDH Mouse Recombinant
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In Stock
Cat. No.
BT8595
Source
E.coli.
Synonyms
Glyceraldehyde-3-phosphate dehydrogenase, GAPDH, Peptidyl-cysteine S-nitrosylase GAPDH, Gapdh, Gapd, Glyceraldehyde-3-phosphate dehydrogenase isoform 2, G3PD, GAPD, HEL-S-162eP.
Appearance
Sterile Filtered colorless solution.
Purity
Greater than 95% as determined by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

Production and Applications

Recombinant GAPDH Mouse is produced in E. coli as a His-tagged protein for experimental use . Key applications include:

ApplicationDetails
Western BlottingLoading control (e.g., Abcam’s mAbcam 9484, 1 µg/mL)
ImmunohistochemistryDetection in frozen tissue sections (1:100 dilution)
Vaccine DevelopmentTarget for Group B Streptococcus (GBS) vaccines in adult mice

Anti-Inflammatory Effects in Sepsis-Related Acute Lung Injury (ALI)

Pre-treatment with GAPDH in a mouse model of lipopolysaccharide (LPS)-induced ALI/ARDS reduced mortality and inflammation :

ParameterPBS Pre-TreatmentGAPDH Pre-Treatment
Survival Rate (48h)0%80%
Serum IL-6 (12h)↑↑↑↓↓↓
Pulmonary NeutrophilsHigh infiltrationMinimal infiltration

Mechanisms:

  • Suppressed proinflammatory cytokines (IL-6, TNF-α, IFN-γ) .

  • Reduced chemokine expression (MIP-2, MIG, RANTES) .

Vaccine Target for Group B Streptococcus (GBS)

A GAPDH-based vaccine showed efficacy in protecting adult mice against GBS infections, highlighting its role in bacterial pathogenesis .

Pseudogene Interference in RT-PCR

Mouse Gapdh has 197 pseudogenes, complicating mRNA quantification . These intronless pseudogenes share high homology with authentic transcripts, leading to false positives in RT-PCR .

GenePseudogenesImplications
Gapdh197Risk of amplifying non-functional transcripts
HPRT1Preferred reference gene due to fewer PGs

Recommendation: Use primers targeting exon-exon junctions or alternative genes (e.g., Hprt) for normalization .

Product Specs

Introduction
Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is an enzyme that plays a crucial role in glycolysis, a metabolic pathway that converts glucose into energy. GAPDH catalyzes the reversible conversion of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate. This enzyme is a tetramer, meaning it consists of four identical subunits, each with a molecular weight of 36 kDa. Beyond its function in glycolysis, GAPDH participates in various cellular processes, including membrane fusion, microtubule organization, phosphotransferase activity, nuclear RNA export, DNA replication, and DNA repair.
Description
This product consists of recombinant mouse GAPDH protein produced in E. coli. It is a single, non-glycosylated polypeptide chain containing 356 amino acids (amino acids 1-333) and has a molecular mass of 38.2 kDa. A 23 amino acid His-tag is fused to the N-terminus of the protein to facilitate purification, which is achieved using proprietary chromatographic techniques.
Physical Appearance
Clear, colorless, and sterile-filtered solution.
Formulation
The GAPDH protein is provided at a concentration of 0.25 mg/ml in a solution of phosphate-buffered saline (pH 7.4) containing 20% glycerol and 1 mM dithiothreitol (DTT).
Stability
For short-term storage (up to 2-4 weeks), the product can be stored at 4°C. For long-term storage, it is recommended to freeze the product at -20°C. The addition of a carrier protein such as bovine serum albumin (BSA) or human serum albumin (HSA) to a final concentration of 0.1% is advised for long-term storage. Repeated freeze-thaw cycles should be avoided to maintain protein stability.
Purity
The purity of the GAPDH protein is greater than 95% as determined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE).
Synonyms
Glyceraldehyde-3-phosphate dehydrogenase, GAPDH, Peptidyl-cysteine S-nitrosylase GAPDH, Gapdh, Gapd, Glyceraldehyde-3-phosphate dehydrogenase isoform 2, G3PD, GAPD, HEL-S-162eP.
Source
E.coli.
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MGSMVKVGVN GFGRIGRLVT RAAICSGKVE IVAINDPFID LNYMVYMFQY DSTHGKFNGT VKAENGKLVI NGKPITIFQE RDPTNIKWGE AGAEYVVEST GVFTTMEKAG AHLKGGAKRV IISAPSADAP MFVMGVNHEK YDNSLKIVSN ASCTTNCLAP LAKVIHDNFG IVEGLMTTVH AITATQKTVD GPSGKLWRDG RGAAQNIIPA STGAAKAVGK VIPELNGKLT GMAFRVPTPN VSVVDLTCRL EKPAKYDDIK KVVKQASEGP LKGILGYTED QVVSCDFNSN SHSSTFDAGA GIALNDNFVK LISWYDNEYG YSNRVVDLMA YMASKE.

Product Science Overview

Structure and Function

GAPDH is a tetrameric protein composed of identical subunits, each with a molecular weight of approximately 37 kDa . The enzyme’s active site contains a cysteine residue that plays a pivotal role in its catalytic mechanism. GAPDH is not only involved in glycolysis but also participates in various cellular processes, including:

  • Apoptosis: GAPDH has been implicated in the regulation of programmed cell death.
  • Gene Expression: It plays a role in the transcriptional regulation of certain genes.
  • Nuclear Transport: GAPDH is involved in the transport of tRNA and other molecules into the nucleus .
Recombinant GAPDH

Recombinant GAPDH, particularly from mouse sources, is commonly used in research due to its high purity and activity. The recombinant form is produced using genetic engineering techniques, where the GAPDH gene from mice is cloned and expressed in a suitable host system, such as E. coli. This allows for the production of large quantities of the enzyme, which can be purified for various experimental applications.

Applications in Research

GAPDH is widely used as a loading control in Western blotting and other protein quantification techniques due to its consistent expression across different tissues and conditions . Its stable expression makes it an ideal reference protein for normalizing the levels of other proteins of interest.

Additionally, GAPDH is used in studies related to:

  • Metabolic Pathways: Understanding the regulation and function of glycolysis.
  • Disease Mechanisms: Investigating its role in diseases such as cancer, neurodegenerative disorders, and diabetes.
  • Drug Development: Screening for potential inhibitors or modulators of GAPDH activity .

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