Recombinant Human MDH1 His-tag Protein, CF

Recombinant Human MDH1 His-tag (Catalog # 11632-MH) is measured by its ability to produce malate from oxaloacetate.

2 μg/lane of Recombinant Human MDH1 His-tag (Catalog # 11632-MH) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 35-38 kDa, under reducing conditions.

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human MDH1 His-tag Protein, CF Summary

• Details of Functionality: Measured by its ability to produce malate from oxaloacetate.
  The specific activity is >55,000 pmol/min/μg, as measured under the described conditions.
• Source: E. coli-derived human MDH1 protein
  Met1-Ala334, with a C-terminal 6-His tag
• Accession #: P40925.4
• N-terminal Sequence: Met1 & Ser2
• Protein/Peptide Type: Recombinant Enzymes
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 37 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.
• SDS-PAGE: 35-38 kDa, under reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris and NaCl.
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 100 mM NaCl, pH 9.0
  • Recombinant Human Malate Dehydrogenase 1 (rhMDH1) (Catalog # 11632-MH)
  • beta -Nicotinamide adenine dinucleotide, reduced disodium salt hydrate (NADH), 20 mM stock in 0.1 M Sodium Borate, pH 9.0
  • Oxaloacetic Acid, 100 mM Stock in deionized water
  • Clear 96-Well Plate
  • Plate Reader with Absorbance Read Capability
  1. Dilute rhMDH1 to 0.5 µg/mL in Assay Buffer.
  2. Prepare a substrate mixture containing 1.6 mM NADH and 2 mM Oxaloacetic Acid in Assay Buffer.
  3. In a plate, load 50 µL of 0.5 µg/mL rhMDH1 and start the reaction by adding 50 µL of substrate mixture. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of substrate mixture.
  4. Read plate at 340 nm (absorbance) in kinetic mode for 10 minutes.
  5. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Vmax* (OD/min) x well volume (L) x 10^12 pmol/mol x (-1)) / (epsilon ** (M^-1cm^-1) x path corr.*** (cm) x amount of enzyme (µg))

  *Adjusted for Substrate Blank
  **Using the extinction coefficient 6220 M^-1cm^-1
  ***Using the path correction of 0.320 cm
  Per Well:
  • rhMDH1: 0.025 µg
  • NADH: 0.8 mM
  • Oxaloacetic Acid: 1 mM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human MDH1 His-tag Protein, CF

• aromatic alpha-keto acid reductase
• cytoplasmic
• Cytosolic malate dehydrogenase
• DEE88
• Diiodophenylpyruvate reductase
• EC 1.1.1.37
• EC 1.1.1.96
• EIEE88
• epididymis secretory protein Li 32
• HEL-S-32
• KAR
• malate dehydrogenase 1, NAD (soluble)
• Malate dehydrogenase, cytoplasmic
• malate dehydrogenase, cytoplasmic; malate dehydrogenase, peroxisomal
• MDH1
• MDHA
• MDH-s
• MGC:1375
• MOR2
• soluble malate dehydrogenase

Background

Malate Dehydrogenase 1 (MDH1), also known as aromatic alpha-keto acid reductase, belongs to the family of nicotinamide adenine dinucleotide (NAD)-dependent dehydrogenases and is a key enzyme in the malate-aspartate shuttle where it plays a critical role in the regulation of metabolic activity in humans (1, 2). MDH1 is an active homodimer of monomers that each contain the characteristic conserved Rossman fold, similar NAD binding sites, and dimeric quaternary structure of MDH enzymes (3). MDH is ubiquitously expressed in cells in two separate isoforms that differ in cellular localization and share only 26% sequence homology (3). While MDH2 is a mitochondrial form involved in the regulation of mitochondrial NAD levels within the citric acid cycle, MDH1, the cytosolic form, plays an important role in the regulation of cytosolic NAD levels. Increased cytosolic NAD levels are necessary for maintaining enhanced glycolysis of proliferating cancer cells (4) and MDH1 is overexpressed in tumors and correlated with poor prognosis (5-8).  In addition, MDH1 has been found to serve as a potential marker in several diseases with inflammation and deficiency of MDH1 is associated with early onset severe encephalopathy and associated with acute liver failure (2, 9-11) making it a metabolic therapeutic target of interest with several potential applications (1, 3, 6, 12, 13).  

1. Godesi, S. et al. (2023) Pharmaceuticals 16:683.
2. Khamis, A.A. et al. (2024). Genes Nutr. 19:20.
3. McCue, W.M and B.C Finzel (2022) ASC Omega 7:207.
4. Lunt, S.Y. et al. (2011) Annu. Rev. Cell Dev. Biol. 27:441.
5. Wang, Y.P. et al. (2016) Mol. Cell. 64:673.
6. Zhang, B. et al. (2017) J. Cancer 8:2088.
7. Sun, W. et al. (2024) BMC Cancer. 24:905.
8. Wang, J. et al. (2024) Clin. Transl. Med. 14:e1680.
9. Broeks, M.H. et al. (2019) Hum. Genet. 138:1247.
10. Shi, C. et al. (2024) iScience. 27:109678.
11. Zeng, X. et al. (2024) Mol. Neurodegener. 19:68.
12. Hanse, E. et al. (2017) Oncogene 36:3915.
13. Naik, R. et al. (2017) J. Med. Chem. 60:8631.

Bioinformatics

• Uniprot:
  • P40925.4()