Glucose Colorimetric Assay Kit II Kit
- Detection Method
- Detection Range
- 0.02-10 mM
- Minimum Detection Limit
- 0.02 mM
- Biochemical Assay (BCA)
- Sample Type
- Cell Culture Supernatant, Plasma, Serum, Tissue Samples
Glucose Assay Kit II provides direct measurement of glucose in various biological samples (e.g., serum, plasma, other body fluids, food, growth media, etc.). In this assay, glucose is specifically oxidized to generate a product which reacts with a dye to generate color (lambda = 450 nm) whose intensity is proportional to glucose concentration. The method is rapid, simple, sensitive, and suitable for high throughput.
This assay is particularly suitable for serum and urine samples since it is unaffected by reducing substances which can interfere with other suppliers offering oxidase-based kits. The assay is also suitable for monitoring glucose level during fermentation and glucose feeding in protein expression processes.
The kit can detect glucose concentrations in the range of 20 ?M - 10 mM.
- Glucose Assay Kit II: High-Throughput Suitable Colorimetric Assay to Measure Glucose in a variety of Biological Samples such as Serum, Plasma, Urine, other body fluids, Food, Fermentation, Growth media, etc. Rapid, Simple & Sensitive.
Glucose Assay Buffer
Glucose Substrate Mix
Glucose Enzyme Mix
Glucose Standard (100 mM)
- Application Notes
- The kit can detect glucose concentrations in the range of 20μM - 10 mM.
Further details regarding sample type: Cell and tissue culture supernatants, urine, plasma and serum, as well as many other biological fluids, fermentation media, food samples etc.
- Reagent Preparation
Glucose Substrate Mix: Dissolve in 220 µL Glucose Assay Buffer. Aliquot and store at - 20 °C, protect from light and moisture. Use within two months.
Glucose Enzyme Mix: Dissolve in 220 µL Glucose Assay Buffer. Aliquot and store at -20 °C. Keep on ice while in use. Use within two months.
- Assay Procedure
Glucose Assay Protocol:
1. Standard Curve Preparations: Dilute the Glucose Standard to 1 nM/µL by adding 10 µL of the Glucose Standard to 990 µL of Glucose Assay Buffer, mix well. Add 0, 2, 4, 6, 8, 10 µL into a series of wells of a 96 well plate. Adjust volume of all wells to 50 µL with Glucose Assay Buffer to generate 0, 2, 4, 6, 8, 10 nM/well of Glucose Standard.
2. Sample Preparations: Prepare test samples in 50 µL/well with Glucose Assay Buffer in a 96-well plate. If using serum sample, serum (0.5-2 µL/assay. Normal serum contains approx. 5 nM/µL glucose) can be directly diluted in the Glucose Assay Buffer. It is recommended to deproteinate samples by centrifugation using a 10 kDa spin column (ABIN413915) to remove enzymes and interfering proteins. For unknown sample, we suggest testing several doses of your sample to make sure the readings are within the standard curve range.
3. Glucose Reaction Mix: Mix enough reagents for the number of assays to be performed: For each well, prepare a total 50 µL Reaction Mix containing: 46 µL Glucose Assay Buffer 2 µL Glucose Enzyme Mix 2 µL Glucose Substrate Mix
4. Mix well. Add 50 µL of the Reaction Mix to each well containing the Glucose Standard and test samples. Mix well.
5. Incubate the reaction for 30 minutes, protect from light.
6. Measure absorbance at 450nm in a microplate reader.
- Calculation of Results
Correct background by subtracting the value derived from the 0 glucose control from all readings (Note: The background reading can be significant and must be subtracted from sample readings). Plot the standard curve. Apply the sample readings to the standard curve. Glucose concentrations of the test samples can then be calculated: C = Sa/Sv (nM/µL or µM/mL, or mM) Where Sa is sample amount (in nmol)calculated from standard curve. Sv is sample volume (in µL) added into the sample wells.
- For Research Use only
- Handling Advice
- Protect from light. Warm the Glucose Assay Buffer to room temperature and briefly centrifuge vials before opening. Read the entire protocol before performing the assay.
- -20 °C
- Expiry Date
- 12 months
Cui, Shi, Xie, Wei, Jia, Zheng, Gao, Huang, Xie: "FOXM1 promotes the warburg effect and pancreatic cancer progression via transactivation of LDHA expression." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 20, Issue 10, pp. 2595-606, 2014 (PubMed).
Shi, Cui, Du, Wei, Jia, Zhang, Zhu, Gao, Xie: "A novel KLF4/LDHA signaling pathway regulates aerobic glycolysis in and progression of pancreatic cancer." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 20, Issue 16, pp. 4370-80, 2014 (PubMed).
Makinoshima, Takita, Matsumoto, Yagishita, Owada, Esumi, Tsuchihara: "Epidermal growth factor receptor (EGFR) signaling regulates global metabolic pathways in EGFR-mutated lung adenocarcinoma." in: The Journal of biological chemistry, Vol. 289, Issue 30, pp. 20813-23, 2014 (PubMed).
Liu, Blouin, Santacruz, Lan, Andriamihaja, Wilkanowicz, Benetti, Tomé, Sanz, Blachier, Davila: "High-protein diet modifies colonic microbiota and luminal environment but not colonocyte metabolism in the rat model: the increased luminal bulk connection." in: American journal of physiology. Gastrointestinal and liver physiology, Vol. 307, Issue 4, pp. G459-70, 2014 (PubMed).
Yu, Zhu, Lin, Zhang, Tian, Hu, Yang: "Functional Analyse of GLUT1 and GLUT12 in Glucose Uptake in Goat Mammary Gland Epithelial Cells." in: PLoS ONE, Vol. 8, Issue 5, pp. e65013, 2013 (PubMed).
Avery, Bumpus: "Valproic acid is a novel activator of AMP-activated protein kinase and decreases liver mass, hepatic fat accumulation, and serum glucose in obese mice." in: Molecular pharmacology, Vol. 85, Issue 1, pp. 1-10, 2013 (PubMed).
Harris, Esain, Frechette, Harris, Cox, Cortes, Garnaas, Carroll, Cutting, Khan, Elks, Renshaw, Dickinson, Chang, Murphy, Paw, Vander Heiden, Goessling, North: "Glucose metabolism impacts the spatiotemporal onset and magnitude of HSC induction in vivo." in: Blood, Vol. 121, Issue 13, pp. 2483-93, 2013 (PubMed).
Paczkowski, Silva, Schoolcraft, Krisher: "Comparative importance of fatty acid beta-oxidation to nuclear maturation, gene expression, and glucose metabolism in mouse, bovine, and porcine cumulus oocyte complexes." in: Biology of reproduction, Vol. 88, Issue 5, pp. 111, 2013 (PubMed).
Sánchez-Solana, Laborda, Baladrón: "Mouse resistin modulates adipogenesis and glucose uptake in 3T3-L1 preadipocytes through the ROR1 receptor." in: Molecular endocrinology (Baltimore, Md.), Vol. 26, Issue 1, pp. 110-27, 2012 (PubMed).
Zhang, Yang, Untereiner, Ju, Wu, Wang: "Hydrogen sulfide impairs glucose utilization and increases gluconeogenesis in hepatocytes." in: Endocrinology, Vol. 154, Issue 1, pp. 114-26, 2012 (PubMed).
Fan, Hitosugi, Chung, Xie, Ge, Gu, Polakiewicz, Chen, Boggon, Lonial, Khuri, Kang, Chen: "Tyrosine phosphorylation of lactate dehydrogenase A is important for NADH/NAD(+) redox homeostasis in cancer cells." in: Molecular and cellular biology, Vol. 31, Issue 24, pp. 4938-50, 2011 (PubMed).
Kuroda, Yamazaki, Abe, Sakimura, Takayanagi, Iwai: "Basic leucine zipper transcription factor, ATF-like (BATF) regulates epigenetically and energetically effector CD8 T-cell differentiation via Sirt1 expression." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 108, Issue 36, pp. 14885-9, 2011 (PubMed).
- Cui, Shi, Xie, Wei, Jia, Zheng, Gao, Huang, Xie: "FOXM1 promotes the warburg effect and pancreatic cancer progression via transactivation of LDHA expression." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 20, Issue 10, pp. 2595-606, 2014 (PubMed).
- Target Type
- Glucose is an important fuel source to generate the universal energy molecule ATP. Serum glucose level is a key diagnostic parameter for many metabolic disorders.