Recombinant Human Lactate Dehydrogenase B Protein

Recombinant Human Lactate Dehydrogenase B Protein Summary

Description	A recombinant protein with a His tag and corresponding to amino acids 1 – 334 of LDHB. Source: E. coli Amino Acid Sequence: MGSSHHHHHH SSGLVPRGSH MATLKEKLIA PVAEEEATVP NNKITVVGVG QVGMACAISI LGKSLADELA LVDVLEDKLK GEMMDLQHGS LFLQTPKIVA DKDYSVTANS KIVVVTAGVR QQEGESRLNL VQRNVNVFKF IIPQIVKYSP DCIIIVVSNP VDILTYVTWK LSGLPKHRVI GSGCNLDSAR FRYLMAEKLG IHPSSCHGWI LGEHGDSSVA VWSGVNVAGV SLQELNPEMG TDNDSENWKE VHKMVVESAY EVIKLKGYTN WAIGLSVADL IESMLKNLSR IHPVSTMVKG MYGIENEVFL SLPCILNARG LTSVINQKLK DDEVAQLKKS ADTLWDIQKD LKDL
Preparation Method	E.coli
Details of Functionality	Specific activity is > 300 units/mg, in which one unit will convert 1.0 umole of pyruvate to L-lactate and beta-NAD per minute at pH 7.5 at 37C.
Protein/Peptide Type	Recombinant Protein
Gene	LDHB
Purity	>95% pure by SDS-PAGE

Applications/Dilutions

Application Notes

The purity of this protein is > 95% by SDS-PAGE. Molecular weight is 38.8 kDa (354aa) confirmed by MALDI-TOF

Assay
1. Prepare a 1.5 ml reaction mix into a suitable container, adjust to pH7.5 at 37C, and pre-chill on ice before use: The final concentrations are 100mM Sodium phosphate, 0.12 mM beta-nicotinamide adenine dinucleotide, reduced form, 2.3mM Pyruvate, 0.033% (w/v) bovine serum albumin.
2. Equilibrate to 37C and monitor at A340nm until the value is constant using a spectrophotometer
3. Add 50ul of recombinant LDHB protein in various concentrations (0.1ug, 0.5ug) in assay buffer.
4. Mix by inversion and record the increase at A340nm for 5 minutes.

Theoretical MW

38.8 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 -80C. Avoid freeze-thaw cycles.
Buffer	20mM Tris-HCl buffer(pH 8.0) containing 10% glycerol, 1mM DTT
Concentration	1.0 mg/ml
Purity	>95% pure by SDS-PAGE

Alternate Names for Recombinant Human Lactate Dehydrogenase B Protein

EC 1.1.1
EC 1.1.1.27
Epididymis Secretory Protein Li 281
HEL-S-281
Lactate Dehydrogenase B
LDH heart subunit
LDHB
LDH-B
LDHBD
LDH-H
L-lactate dehydrogenase B chain
Renal Carcinoma Antigen NY-REN-46
TRG-5

Background

LDHB, also known as L-lactate dehydrogenase B chain, is a member of the lactate dehydrogenase family. It is an oxidoreductase which catalyses the interconversion of pyruvate and lactate with concomitant interconversion of NADH and NAD+. As this protein can also catalyze the oxidation of hydroxybutyrate, it is occasionally called Hydroxybutyrate Dehydrogenase (HBD). The LDH family consists of three members, LDH-A, LDH-B and LDH-C. LDHs function as powerful markers for germ cell tumors. Recombinant human LDHB protein, fused to His-tagging at N-terminus, was expressed in E.coli and purified by using conventional chromatography techniques.

PMID: 24012368