Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Adherent cells, Serum, Suspension cells, Tissue, Tissue Extracts
- Sensitivity: 0.1 mU/well
Product nameHexokinase Activity Assay Kit (Colorimetric)
See all Hexokinase kits
Sample typeSerum, Tissue, Adherent cells, Suspension cells, Tissue Extracts
Assay typeEnzyme activity (quantitative)
Sensitivity< 0.1 mU/well
Assay time1h 00m
Species reactivityReacts with: Mammals, Other species
Hexokinase Activity Assay Kit (ab136957) uses a simple, sensitive and rapid procedure which can detect hexokinase activity to below 0.1 mU/well.
In the hexokinase assay protocol, glucose is converted to glucose-6-phosphate by hexokinase. The glucose-6-phosphate is then oxidized by glucose-6-phosphate dehydrogenase to form NADH. The NADH reduces a colorless probe to a colored product with strong absorbance at 450 nm.
Hexokinase activity assay protocol summary:
- add samples and standards to wells
- add reaction mix and incubate for 20-60 min
- analyze repeatedly with a microplate reader during the incubation
Hexokinases (HK) are found in many organisms including bacteria, plants and mammals and play an important role in glucose metabolism. Hexokinases phosphorylate glucose and generate glucose-6-phosphate for glycolysis. Hexokinases have four isoforms (HK-I, II, III and IV). HK-I, HK-II and HK-III have low Km, while HK-IV has 100 fold high Km.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 100 tests Assay Buffer WM 1 x 25ml Developer (Lyophilized) Red 1 vial Hexokinase Coenzyme Purple 1 vial Hexokinase Enzyme Mix Green 1 vial Hexokinase Substrate White 1 x 1ml NADH Standard ( lyophilized) Yellow 1 vial Positive Control Orange 1 vial
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
Hexokinase Activity measured in cell lysates showing quantity (nmol) per 1 mln cells after 35 min of incubation.
Samples with the concentration of 5e6 cells/mL were used. Samples were diluted 1-27 fold.
Hexokinase Activity measured in tissue lysates showing quantity (nmol) per mg of extracted protein after 35 min of incubation.
Protein concentration for samples varied from 13 mg/mL to 43 mg/mL. Samples were diluted 1-9 fold.
Standard curve: mean of duplicates (+/- SD) with background reads subtracted
NADH standard curve (a). Hexokinase activity in Positive Control and Jurkat cell lysate (40 µg) (b).
In this kit glucose is converted to glucose-6-phosphate by hexokinase; the glucose-6-phosphate is oxidized by glucose-6-phosphate dehydrogenase to form NADH, which reduces a colorless probe to a colored product with strong absorbance at 450 nm.
ab136957 has been referenced in 8 publications.
- Iftikhar A et al. Effect of Caesalpinia bonduc Polyphenol Extract on Alloxan-Induced Diabetic Rats in Attenuating Hyperglycemia by Upregulating Insulin Secretion and Inhibiting JNK Signaling Pathway. Oxid Med Cell Longev 2020:9020219 (2020). PubMed: 32256963
- Luo H et al. Apelin-13 Suppresses Neuroinflammation Against Cognitive Deficit in a Streptozotocin-Induced Rat Model of Alzheimer's Disease Through Activation of BDNF-TrkB Signaling Pathway. Front Pharmacol 10:395 (2019). PubMed: 31040784
- Yao J et al. By blocking hexokinase-2 phosphorylation, limonin suppresses tumor glycolysis and induces cell apoptosis in hepatocellular carcinoma. Onco Targets Ther 11:3793-3803 (2018). PubMed: 30013360
- Bramer SA et al. Hexokinase 2 drives glycogen accumulation in equine endometrium at day 12 of diestrus and pregnancy. Reprod Biol Endocrinol 15:4 (2017). Functional Studies ; Horse . PubMed: 28056994
- Kim DJ et al. Increased glucose metabolism and alpha-glucosidase inhibition in Cordyceps militaris water extract-treated HepG2 cells. Nutr Res Pract 11:180-189 (2017). PubMed: 28584574
- Albert V et al. mTORC2 sustains thermogenesis via Akt-induced glucose uptake and glycolysis in brown adipose tissue. EMBO Mol Med 8:232-46 (2016). PubMed: 26772600
- Maiso P et al. Metabolic signature identifies novel targets for drug resistance in multiple myeloma. Cancer Res 75:2071-82 (2015). PubMed: 25769724
- Hamabe A et al. Combined evaluation of hexokinase 2 and phosphorylated pyruvate dehydrogenase-E1a in invasive front lesions of colorectal tumors predicts cancer metabolism and patient prognosis. Cancer Sci 105:1100-8 (2014). PubMed: 25060325