About Calzyme Contact Us Locations Our Policies

  Welcome to Calzyme!

Glycerol-3-Phosphate Dehydrogenase (G3PDH)

(L-glycerol-3-phosphate:NAD+ oxidoreductase; EC 1.1.1.8) Glycerol-3-phosphate dehydrogenase (GPDH) catalyzes the following reaction: GPDH Dihydroxyacetone phosphate + NADH + H+ ------> L-glycerol-3-phosphate + NAD+ GPDH activity has been demonstrated in a wide variety of species belonging to animal as well as plant kingdoms (Lin, E.C.C., Ann. Rev. Biochem, 46, 765, 1977). A marked reduction in GPDH activity was observed in chronic plaques of multiple sclerosis (MS) (Hirsch, H.E., Blanco, C.E. and M.E. Parks, J. Neurochem., 34(3), 760, 1980). This may be of diagnostic and clinical significance. GPDH can also be used for glycerol determination in triglyceride kits (Sugiura, M., Oikawa, R., Hirano, K., Maeda, H., Yoshimura, H., Sugiyama, M. and T. Kuratsu, Clin Chem Acta, 81, 125, 1977).




The amount of enzyme which will convert one micromole of dihydroxyacetone phosphate to alpha-glycerophosphate per minute at pH 7.9 at 25C.


The rate of decrease in the absorbancy at 340 nm, resulting from the oxidation of NADH, is a measure of GPDH activity.


  1. 0.05 M Triethanolamine hydrochloride buffer, pH 7.9.
  2. 0.006 M NADH disodium salt (3.9 mg/ml) in buffer. ,LI> 0.024 M Dihydroxyacetone phosphate, dimethylketal dicyclohexylamine salt (10 mg/ml). Dissolve 250 mg dihydroxyacetone phosphate, dimethylketal dicyclohexylamine salt in about 8 ml distilled water. Add Dowex-50 W, hydrogen form, until pH reaches approximately 2. Filter, wash and incubate filtrate at 40C for 4-6 hours. Cool and adjust pH to 4.5 with 0.4 M sodium carbonate. Bring the volume up to 25 ml with distilled water. Store frozen in 1 ml aliquots (substrate).
  3. Glycerol-3-phosphate dehydrogenase (GPDH) solution (10 mg/ml). Dilute further, using 1% cold bovine serum albumin to give a final concentration of 0.5-1.0 U/ml. Must be prepared fresh immediately prior to assay.



  1. Set spectrophotometer (equipped with strip chart recorder and temperature control) at 340 nm and 25C.
  2. Into a cuvette pipette the following reagents:
    Triethanolamine HCl buffer         2.7 ml 
    NADH                               0.1 ml 
    Dihydroxyacetone phosphate         0.1 ml 
    
  3. Mix and incubate at 25C for 5 minutes in the spectrophotometer, to attain temperature equilibration.
  4. Record blank rate, if any, at 340 nm for 2-3 minutes.
  5. Initiate the reaction by adding 0.1 ml of fresh GPDH (enzyme) solution to the cuvette. Monitor the reaction for 5-10 minutes at 340 nm.
  6. Calculate (delta)E340 nm/min

 

©1999-2017 Calzyme All Rights Reserved powered by corezon
All logos and trademarks are properties of their respective companies. Product price, specification, warranty and availability subject to change without notice.
2-Phosphoenol Pyruvate Acid 3-Indoxyl Phosphate
3-Phosphoglycerate Kinase Acetone Powders
Acetyl Cholinesterase Acid Phosphatase
Actin Acylase
Adenosine 5'-diphosphate Adenosine 5'-monophosphate (AMP)
Adenosine 5'-triphosphate (ATP) Adenosine Deaminase
Alcohol Dehydrogenase Aldehyde Dehydrogenase
Aldolase Alkaline Phosphatase
alpha-Amylase alpha-D-Glucose-1-Phosphate
alpha-Glucosidase alpha-Ketoglutarate
Amylopectin Amylose
Apo-d-Amino Acid Oxidase Ascorbate Oxidase
Avidin beta-Amylase
beta-Glucosidase beta-Glucronidase
beta-NAD beta-NADH
beta-NADPH Bromelain
Butyryl Cholinesterase C-Reactive Protein
Carbonic Anhydrase Carboxypeptidase A
Carboxypeptidase B Carboxypeptidase Y
Catalase Catechol-o-Methyl Transferase
Cathepsin D Cholesterol Esterase
Chymotrypsin d-Amino Acid Oxidase
d-Glucose-6-Phosphate Deoxyribonuclease (DNase)
Deoxyribonucleic Acid (DNA) Elastase
Enterokinase Ferritin
G-6-phosphate Dehydrogenase gamma-Glutamyl Transferase
Glucose Oxidase Glutamate Dehydrogenase
Glutathione Peroxidase Glycerol Kinase
Glycerol-3-phosphate Dehydrogenase Glycosylated Albumin
Glycosylated Hemoglobin GOT (AST)
GPT (ALT) Hemoglobin
 Hexokinase Histamine-n-Methyl Transferase
Hyaluronic Acid Hyaluronidase
Insulin L-Arginase
L-gamma-Glutamide Nitroanilide Lactate Dehydrogenase
Lactoperoxidase Leucine Aminopeptidase
Lipase Lipoamide Dehydrogenase
Lipoxidase Luciferase
Lysozyme Malate Dehydrogenase
Mutarotase Myelin
Myoglobin Myosin
Oxyhemoglobin p-Nitrophenyl Phosphate
Pepsin Peroxidase
Phosphorylase-b Phspho(enol) Pyruvate Carboxylase
PNMT Prophospholipase A2
Protein Methylase II Pyruvate Kinase
Renin Rennin
Ribonuclease Sedoheptulose
Stachyose Superoxide Dismutase
Thyroglobulin Triose Phosphate Isomerase
Trypsin Trypsin Inhibitor
Tyrosinase Urease
Uricase Xanthine Oxidase


abc