EnzyChrom™ D-Amino Acid Assay Kit

-20 °C

Will this kit detect L-amino acids?
The DAA enzyme is specific to D-amino acids only.

What samples have you tested?
This kit has been tested on rat brain tissue, mouse brain tissue, cerebrospinal fluid, and dairy products like whole milk.

Can I store unused reagents for future use?
Yes, unused reagents can be stored according to the assay protocol.

How do you use this kit on cerebrospinal fluid?
CSF can be assayed directly and an internal standard should be used.
Samples will need three separate reactions:
1) sample plus standard
2) sample alone
3) sample blank.
For the internal standard prepare 200 uL 1250 uM standard by mixing 125 uL 2 mM Standard and 75 uL dH2O.
For the sample plus standard well, add 5 uL 1250 uM standard and 20 uL sample.
For the sample and sample blank wells, add 5 uL dH₂O and 20 uL sample.
Prepare enough Working Reagent (WR) for all assay wells by mixing, for each well, 85 uL Assay Buffer, 1 uL DAA Enzyme, 1 uL HRP Enzyme, 1 uL Dye Reagent. Prepare a WR without the DAA Enzyme for the sample blank well(s).
Add 80 uL of the WR to each well. Tap plate briefly to mix.
Incubate at room temperature for 60 min. Use a plate reader to read OD_570nm.
The sample D-amino acid concentration is computed as follows:

where R_SAMPLE, R_BLANK, and R_STANDARD are the OD or fluorescence values of the Sample, Sample Blank, and the Sample plus Standard respectively. n is the sample dilution factor. Note: The volume of the internal standard is 4x lower than the sample volume; thus, the internal standard concentration should be divided by 4.


For more detailed product information and questions, please feel free to send an inquiry to info@hoelzel.de

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