The standard curve should be plotted after subtracting the 0 Standard reading from all Standard readings. Additionally, the PFK assay buffer suggested for sample preparation can be adjusted proportionally. For instance, 100 µl of PFK assay buffer can be used for 10 mg of tissue or 1 x 10^6 cells.
MAK093
Phosphofructokinase (PFK) Activity Colorimetric Assay Kit
sufficient for 100 colorimetric tests
Synonym(s):
PFK Activity Assay Kit
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About This Item
UNSPSC Code:
12161503
NACRES:
NA.84
Pricing and availability is not currently available.
Recommended Products
usage
sufficient for 100 colorimetric tests
detection method
colorimetric
relevant disease(s)
cancer; orthopedic diseases; aging/geriatric diseases
storage temp.
−20°C
Gene Information
human ... PFKL(5211) , PFKM(5213) , PFKP(5214)
mouse ... PFKL(18641) , PFKM(18642) , PFKP(56421)
rat ... PFKL(25741) , PFKM(65152) , PFKP(60416)
General description
Phosphofructokinase (PFK) catalyzes the third step of glycolysis, the conversion of fructose-6-phosphate to fructose-1,6-diphosphate, the rate limiting step of glycolysis. PFK activity is highly regulated by multiple cofactors in addition to post-translational modifications and can be used to measure glycolytic flux in tissues. Deficiencies in PFK activity can result in the glycogen storage disease, glycogenosis type VII (Tarui′s disease), which is a glycogen storage disease.[1][2]
Application
Suitability
Suitable for the measurement of phosphofructokinase activity, analysis of glucose metabolism and cell signaling and screening anti-cancer drugs in animal tissues (liver, brain, heart, muscles etc.) and cell culture (adherent or suspension cells).
Principle
The Phosphofructokinase Colorimetric Assay kit provides a simple and direct procedure for measuring PFK activity in a variety of samples. PFK activity is determined by a coupled enzyme assay, in which fructose-6-phosphate and ATP is converted to fructose-1,6-diphosphate and ADP by PFK. The ADP is converted by the enzyme mix to AMP and NADH. The resulting NADH reduces a colorless probe resulting in a colorimetric (450 nm) product proportional to the PFK activity present. One unit of PFK is the amount of enzyme that will generate 1.0 μmole of NADH per minute at pH 7.4 at 37 °C.
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Resp. Sens. 1
Storage Class Code
10 - Combustible liquids
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Neng Tang et al.
Biochemical and biophysical research communications, 526(1), 1-7 (2020-03-21)
The mechanisms underlying the initiation and proliferation of liver regeneration (LR) has been extensively studied using the partial hepatectomy (PHx) model, while little is known about the termination of LR. PP2Acα (protein phosphatase 2 A catalytic subunit α isoform) is
Kai Zhou et al.
Cell death & disease, 9(10), 988-988 (2018-09-27)
Plastic phenotype convention between glioma stem cells (GSCs) and non-stem tumor cells (NSTCs) significantly fuels glioblastoma heterogeneity that causes therapeutic failure. Recent progressions indicate that glucose metabolic reprogramming could drive cell fates. However, the metabolic pattern of GSCs and NSTCs
Manipulation of a glycolytic regulator alters growth and carbon partitioning in the marine diatom Thalassiosira pseudonana.
Abbriano R, et al.
Algal research, 32, 250-258 (2018)
Emilio Bueno et al.
Nature microbiology, 3(12), 1346-1353 (2018-10-03)
To survive and proliferate in the absence of oxygen, many enteric pathogens can undergo anaerobic respiration within the host by using nitrate (NO3-) as an electron acceptor1,2. In these bacteria, NO3- is typically reduced by a nitrate reductase to nitrite
D S Matassa et al.
Cell death and differentiation, 23(9), 1542-1554 (2016-05-21)
Tumour cells have long been considered defective in mitochondrial respiration and mostly dependent on glycolytic metabolism. However, this assumption is currently challenged by several lines of evidence in a growing number of tumours. Ovarian cancer (OC) is one of the
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Is the standard curve created from the difference between the final and initial A_450 standard values, given the standards at final and initial time points? Alternatively, is the curve generated solely from the final A_450 standard values? Also, if we have fewer than 2x10^6 cells, how much assay buffer should be used for homogenization when the instructions state to homogenize 2x10^6 cells in 200uL of assay buffer?
1 answer-
Helpful?
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Does this kit work for all isoforms of Phosphofructokinase, such as PFK-M (muscle), PFK-L (liver), and PFK-P (platelet)?
1 answer-
MAK093 is designed based on the indirect detection of phosphofructokinase activity. The ADP generated from the conversion of fructose-6-phosphate to fructose-diphosphate should react with a substrate and be converted to AMP and NADH. This process reduces a colorless probe to a colored product with strong absorbance at 450 nm. Therefore, considering that all three major isoforms in mammals, PFK-M (muscle), PFK-L (liver), and PFK-P (platelet), should catalyze the same enzymatic reaction producing ADP, it is assumed that this kit will not be selective for one specific isoform.
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Does the Positive Control in MAK093 include PFK-P, -L, or -M?
1 answer-
PFK-P (platelets), L (Liver), and M (Muscle) are all mammalian, and since the product is from bacillus sps, the question doesn't really apply to the PFK in use.
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