GAPDH Mouse, Active
GAPDH Mouse Recombinant, Active
Cat. No.
BT8654
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
GAPDH Mouse Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 356 amino acids (1-333a.a.) and having a molecular mass of 38.2kDa.
GAPDH is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.
GAPDH is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.
Product Specs
Introduction
Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is an enzyme that plays a crucial role in glycolysis, a metabolic pathway that breaks down glucose to produce energy. GAPDH is a tetrameric enzyme, meaning it consists of four identical subunits, each with a molecular weight of 36 kDa. This enzyme catalyzes the reversible conversion of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate, a key step in glycolysis. Beyond its role in energy production, 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 the recombinant form of mouse GAPDH, expressed in E. coli. It is a single, non-glycosylated polypeptide chain composed of 356 amino acids (residues 1-333), resulting in a molecular weight of 38.2 kDa. A 23 amino acid His-tag is fused to the N-terminus to facilitate purification, which is achieved through proprietary chromatographic methods.
Physical Appearance
Clear, colorless solution, sterile-filtered.
Formulation
The GAPDH protein is provided in a solution at a concentration of 0.25 mg/ml. The solution is buffered with phosphate-buffered saline (pH 7.4) and contains 20% glycerol and 1 mM dithiothreitol (DTT) as stabilizing agents.
Stability
For short-term storage (up to 4 weeks), the product can be stored at 4°C. For extended storage, it is recommended to freeze the product at -20°C. Adding 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 freezing and thawing should be avoided to maintain protein integrity.
Purity
The purity of this product is greater than 95%, as determined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE).
Biological Activity
This enzyme exhibits a specific activity greater than 40 units/mg. One unit of activity is defined as the amount of enzyme required to catalyze the conversion of 1.0 micromole of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate per minute at a pH of 8.5 and a temperature of 37°C.
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 Expression
GAPDH is a tetrameric enzyme composed of four identical subunits, each with a molecular weight of approximately 36 kDa . The recombinant mouse GAPDH protein is typically expressed in Escherichia coli and purified to a high degree of purity (>95%) using conventional chromatography techniques . The protein sequence includes a His tag at the N-terminus to facilitate purification .
Function and Activity
GAPDH plays a dual role in cellular metabolism:
- Glycolysis: It catalyzes the reversible conversion of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate in the presence of NADPH . This reaction is a key step in the glycolytic pathway, which is crucial for ATP production.
- Nuclear Functions: Beyond its role in glycolysis, GAPDH also has nitrosylase activity, which allows it to participate in various nuclear events such as transcription, RNA transport, DNA replication, and apoptosis .
The specific activity of the recombinant mouse GAPDH is greater than 40 units/mg, defined as the amount of enzyme that converts 1.0 µmole of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate per minute at pH 8.5 at 37°C .
Applications in Research
Due to its critical role in both glycolysis and nuclear functions, GAPDH is a valuable tool in various research applications:
- SDS-PAGE: Used as a loading control in protein electrophoresis to ensure equal protein loading across samples .
- Functional Studies: Investigating the enzyme’s activity and its role in cellular metabolism and nuclear functions .
- Mass Spectrometry (MS): Analyzing protein interactions and modifications .
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