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Aldolase Activity Colorimetric Assay Kit

Catalog #: K665

In stock

$525.00

Product Details

Cat # +Size K665-100
Size 100 assays
Detection Method Absorbance (OD 450 nm)
Applications Measurement of Aldolase activity in various tissues/cells
Analysis of glycolysis and gluconeogenesis pathways
Features & Benefits • Rapid, convenient and high-throughput adaptable
• Kit can detect aldolase activity less than 0.1 mU
Kit Components • Aldolase Assay Buffer
• Aldolase Substrate (Lyophilized)
• Aldolase Enzyme Mix (Lyophilized)
• Aldolase Developer (Lyophilized)
• NADH Standard (Lyophilized)
• Aldolase Positive Control (Lyophilized)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Aldolase (Fructose-Bisphosphate Aldolase: EC 4.1.2.13) is an important enzyme for both glycolysis and gluconeogenesis. It catalyzes the reversible reaction of fructose-1,6-bisphosphate to glyceraldehyde-3-phosphate & dihydroxyacetone. There are 2 classes of Aldolase - class l: found in animal and plant tissues and class II: found in prokaryotes and lower eukaryotes. Class I Aldolase has 3 isozymes- Type A: found in muscle and red blood cells, Type B: found in liver and kidney and Type C: found in brain. Aldolase A deficiency leads to myopathy & hemolytic anemia. Muscle disease and liver injury can also cause increased serum aldolase. Accurate detection of aldolase activity is valuable for diagnostic and mechanistic studies. In BioVision’s Aldolase Activity Assay, aldolase converts fructose-1,6-bisphosphate to glyceraldehyde-3-phosphate and dihydroxyacetone, and through a series of reactions, reduces a nearly colorless probe to a colored product with absorbance at 450 nm. This assay kit is simple, sensitive, and high-throughput adaptable. Detection limit: less than 0.1 mU of aldolase activity in a variety of samples.


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Will Glyceraldehyde -3-P interfere in this assay?
Yes, endogenous Glyceraldehyde -3-P can interfere with this assay. A parallel background control without the aldolase substrate will helpaccount for this interference.
Does this kit detect all isoforms of Aldolase?
Yes, this kit detects Aldolase based on actvity and hence all Class I and II Aldolases will be detected based on the sample.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Is it possible to use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. It is recommended to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is strongly recommended to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
Can individual components of this kit be purchased separately?
Yes, any of the kit's components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.