Assay Kit Name:
Acetate Colorimetric Assay Kit
• Detection method - Absorbance (450 nm)
• Application - The kit can detect less than 20 μM of Acetate in various samples.
• Animal tissues - Liver, kidney, muscle, heart etc
• Cell culture - Adherent or suspension cells
Features & Benefits:
• Rapid, convenient and sensitive
• Kit contains all necessary reagents for accurate measurement of Acetate levels
• Acetate Assay Buffer
• Acetate Enzyme Mix (Lyophilized)
• ATP (Lyophilized)
• Acetate Substrate Mix (Lyophilized)
• Probe (Lyophilized)
• Acetate Standard (Lyophilized)
Acetate is an important component for biosynthesis. In living organisms, acetylation/deacetylation of proteins is critical for post-translational regulation of their functions. When bound to CoA, Acetyl-CoA is central to metabolism of carbohydrates and fats. It is also involved in the synthesis of biogenic compounds such as neurotransmitter acetylcholine. In Biovision’s Acetate Assay Kit, Acetate is converted to an intermediate in the presence of Acetate Enzyme Mix & Acetate Substrate Mix. The intermediate reduces a colorless probe to a colored product with strong absorbance at 450 nm. The Acetate Colorimetric Assay Kit is rapid, simple & sensitive. It can detect less than 20 µM Acetate in various samples.
USAGE: For Research Use Only! Not For Use in Humans.
Will Acetoacetate react in this assay?
The Acetate enzyme mix is specific for acetate and will not work on acetoacetate. If you wish to quantify acetoacetate in your samples I would suggest looking at our kit K650 (http://www.biovision.com/acetoacetate-colorimetric-assay-kit-7475.html)
We see very high background values and hence the numbers are negative after subtraction. What is the solution?
Acetate vapors absorbed into the reagents can cause high background. Absorption of vapors is a common issue and this can destabilize the kit components. Special care needs to be taken to prevent this.
Is EDTA usable for blood collection for this assay?
One or more of the enzymes in the kit requires divalent metal cations. Hence EDTA in samples is not advisable for this assay.
Does the kit measure free or bound acetate?
The kit can be used to measure free Acetate in biological samples. If the acetyl group is bound to protein or any other chemical moiety while in solution, it cannot be measured.
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.
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.