Acetyl-CoA Assay Kit

• Cat.No.: Kit-0036
• Product Overview: Acetyl-CoA Assay Kit is used for determining acetyl CoA level in a variety of samples.

Specification

• Storage: Store kit at -20°C, protect from light. Warm Acetyl CoA Assay Buffer to room temperature prior to using it. Briefly centrifuge all small vials prior to opening.

Reviews

01/20/2023

Their products and services are ALWAYS of high standard.

12/01/2022

Detection is very precise. And very easy to use.

Q&As

Is your PicoProbe kit sensitive enough to detect AcetylCoA in HEK293 cytosol?

This assay candetect 10 to 1000 pmol of Acetyl CoA (with detection limit ˜0.4 μM). If you are able to optimize the amount of lysate to obtain Acetyl CoA levels within the detection limit this product should detect it in HEK 293.

Can this protocol be modified to detect malonyl-CoA?

This kit has been specifically designed to detect acetyl CoA and not for malonyl-CoA.

My sample itself is colored, will this affect the fluorescence detection?

Sample color does not interfere with fluorescence detection.

If my sample contains other CoA compounds, will that influence the result of the assay?

Only free CoA will initially be quenched in the first step. Then only the Acetyl CoA will get converted to CoA due to the specific enzymes used. So there should be very minimal interference by any other CoAs.

Can I use bead beater to lysis instead of using the protocol you provide on the website?

As long as you can use the deproteinization protocol along with the bead beater, it should be fine.

I don't have a fluorometer, is it possible to use a spectrophotometer?

Sorry, the spectrophotometer cannot be used

Can neutralized samples be frozen?

It is not the best case scenario to freeze samples repeatedly before performing the assay. However, if you need to freeze the samples, it would be possible after separating the sample from the KClO4 pellet.

Can mitochondrial samples isolated from cultured cells be tested with this kit?

Mitochonrial lysate can be assayed by Kit-0036.we recommended to perform a pilot experiment with a range of dilutions of the sample to determine the optimal concentration/dilution of sample which gives the best results in the linear range of the standard curve.

Is this kit suitable for rat frozen tissues?

This kit can be used with frozen rat tissues.

Can the prepared samples be stored at -80 for experiments later?

The sample can be stored at -80 degree centigrade once prepared. However, we recommend to use the sample immediately after preparation for maximum efficiency.

What is the mechanism of the kit to detect Acetyl-CoA?

The assay measures relative amounts of acetyl CoA in samples indirectly, compared to a standard included with the kit, by converting the acetyl CoA to CoA, and then using the resulting CoA to reduce NAD to NADH, which in turn reacts with the probe to generate a fluorescent signal.

Is picoprobe specific to NADH? Or does it respond positively to other cofactors as well?

In this particular assay the probe is specific to just the NADH.

For the deproteinization step, could perchloric acid method be substituted by using a 10kD MW cut-off spin column?

The deproteinization approach, though not as sensitive as the sample prep, would still benefit from using PCA rather than the 10 kDa spin filters. But you can use the spin filters if you do not have the option of using PCA.

Is deproteinization also necessary for cells in culture?

Deproteinization is recommended for cells in culture.

Do I absolutely need to follow the protocol and use specific reagents?

Yes it is very crucial to do the sample prep exactly as recommended as Acetyl CoA is very sensitive to degradation and you risk the chance of inefficient results if you make changes to the protocol.

Can CoA be measured alone?

In principle, eliminating the quenching and conversion step will allow you to measure CoA alone. However, the kit was not produced nor validated for this purpose, so we cannot guarantee that it will work, nor do we have a CoA standard that has been validated with this method to provide with the kit.