Mdh, Recombinant, E.coli, aa1-312, His-Tag(Malate Dehydrogenase)

Blue Ice

34, 9

Supplied as a liquid in 20mM Tris-HCl, pH 8.0, 1mM DTT, 50mM sodium chloride, 10% glycerol.

Malate dehydrogenase (mdh) belongs to the LDH/MDH superfamily and MDH type 1 family. This enzyme catalyzes the conversion of malate into oxaloacetate (using NAD+) and vice versa (this is a reversible reaction). Malate dehydrogenase is also involved in gluconeogenesis, the synthesis of glucose from smaller molecules.

Source:
Recombinant protein corresponding to aa1-312 from E. coli Mdh, fused to His-tag at N-terminal, expressed in E. coli. Accession: NP_417703.

Molecular Weight:
~34.9kD (336aa)

Specific Activity:
>1000units/mg, and is defined as the amount of enzyme that cleaves 1umole of oxalacetate and beta-NADH to L-malate and beta- NAD/minute at pH 8.0 at 37°C.

Activity Assay:
1. Prepare a 180ul assay buffer into a suitable container. The concentrations are 100mM potassium phosphate (pH8.0), 0.5mM beta-nicotinamide adenine dinucleotide, reduced form, 0.5mM oxalacetic acid
2. Equilibrate to 37°C and monitor at A340nm until the value is constant using a spectrophotometer.
3. Add 20ul of recombinant MDH protein 0.25ug/ml in assay buffer.
4. Mix by inversion and record the decrease at A340nm for 5 minutes.

Amino Acid Sequence:
MGSSHHHHHH SSGLVPRGSH MGSHMKVAVL GAAGGIGQAL ALLLKTQLPS GSELSLYDIA PVTPGVAVDL SHIPTAVKIK GFSGEDATPA LEGADVVLIS AGVARKPGMD RSDLFNVNAG IVKNLVQQVA KTCPKACIGI ITNPVNTTVA IAAEVLKKAG VYDKNKLFGV TTLDIIRSNT FVAELKGKQP GEVEVPVIGG HSGVTILPLL SQVPGVSFTE QEVADLTKRI QNAGTEVVEA KAGGGSATLS MGQAAARFGL SLVRALQGEQ GVVECAYVEG DGQYARFFSQ PLLLGKNGVE ERKSIGTLSA FEQNALEGML DTLKKDIALG EEFVNK

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.