About Calzyme Contact Us Locations Our Policies

  Welcome to Calzyme!

Pyruvate Kinase (PK)

(ATP:Pyruvate 2-0-phosphotransferase; EC 2.7.1.40)

Pyruvate kinase (PK) catalyzes the following reaction:

                               PK
Phospho(enol) pyruvate + ADP ------> Pyruvate + ATP 
Pyruvate kinase is an important enzyme in glucose metabolism. Mammalian PK of various tissues have distinct characteristics which are related to the specific metabolic requirements of each tissue. PK, prepared from rabbit muscle, is extensively used in the quantitative determination of ADP and of enzymes that catalyze the formation of ADP. It can also be used for the determination of phospho(enol) pyruvate and 2-phosphoglycerate. The assay of PK activity in red blood cells has been described by Tanphaichitr and Van Eys (Tanphaichitr, V.S. and Van Eys, J., Clin. Chim. Acta, 41, 41, 1972).


1.Source: Rabbit Muscle
   Form: Freeze-dried 
   Solubility: Soluble in distilled water or dilute buffer 
   Stability: -20 C; -4 F 
   Activity: 250 U/mg protein 
   Protein: 90-95% (biuret) 
   Contaminants: Myokinase <0.005% Glycerol-phosphate dehydrogenase <0.001% Creatine phosphokinase <0.001% Adenosine triphosphatase <0.001% NADH oxidase <0.001% Phospho-glycerate-mutase <0.001% 
   Catalog No.: 107A0250 
$2.00/KU


The amount of enzyme which produces one micromole of pyruvate from phospho(enol)pyruvate, in the presence of ADP, per minute at pH 7.5 and 25°C.


The method of assay has been described by Tietz and Ochoa (Tietz and Ochoa, S., Arch. Biochim. Biophys. Acta., 78, 477, 1958). The rate of decrease in the absorbancy at 340 nm, resulting from the oxidation of NADH, is proportional to the PK activity.


  1. 0.06 M Tris-HCl buffer, pH 7.5, in distilled water.
  2. 0.12 M Magnesium chloride, in distilled water.
  3. 2.25 M Potassium chloride, in distilled water.
  4. 0.006 M Adenosine diphosphate (ADP), disodium, in buffer. Prepare fresh.
  5. 0.043 M Phospho(enol)pyruvate (PEP), trisodium in buffer. Prepare fresh.
  6. 0.006 M NADH, disodium salt, in buffer. Prepare fresh.
  7. Lactate dehydrogenase (LDH) solution, 40 U/ml in buffer. Prepare fresh.
  8. Pyruvate kinase (PK) solution. Using cold 1% bovine serum albumin as solvent, dilute to a final concentration of 0.1-0.2 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 25°C.
  2. Into a cuvette, pipette the following reagents as indicated:
    Tris-HCl buffer, pH 7.5 2.30 ml
    Magnesium chloride 0.10 ml
    Potassium chloride 0.10 ml
    ADP 0.10 ml
    PEP 0.10 ml
    NADH 0.10 ml
    LDH 0.10 ml

    Mix and incubate in spectrophotometer at 25°C for 5 minutes to achieve temperature equilibration.

  3. Determine blank rate, at 340 nm, if any.
  4. Initiate the reaction by adding 0.1 ml of enzyme (PK) solution (0.1-0.2 U/ml) to the cuvette. Monitor the absorbance change at 340 nm for 5 minutes and then calculate µE340 nm/min



Activity (U/mg) = (ΔE340nm/min)(Total Vol.)(Enz. Diln.)
(6.22)(Enz. Vol.)(mg Enz./ml)
 

©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