EnzyChrom™ D-Amino Acid Assay Kit
- For quantitative determination of D-Amino Acids in biological and other samples.
- Fast and sensitive. Linear detection range: 0.86 to 500 µM (colorimetric assay) and 0.18 to 50 µM (fluorimetric assay) for 60 min reaction.
- Convenient. The procedure involves adding a single working reagent and reading after 60 minutes.
- High-throughput. Homogeneous “mix-incubate-measure” type assay. Can be readily automated to process thousands of samples per day.
- OD570nm or FL530/585nm
- Tissue, milk, and other biological samples
- 60 min
- 100 tests
- 0.86 µM (colorimetric assay), 0.18 µM (fluorimetric assay)
- 6 months
More DetailsD-AMINO-ACIDS are not as widespread as their enantiomeric counterparts in proteins but they can be found in organisms ranging from bacteria (cell walls and antibiotics) to mammals (central nervous systems). The presence of D-amino acids in food is also of considerable interest. Racemization of L-amino acids during food processing may affect food quality and nutritional value.BioAssay Systems’ D-amino acid assay uses an enzyme-catalyzed oxidation of D-amino acids to convert a dye into a colored and fluorescent form. The absorbance at 570 nm or fluorescence intensity at λex/em = 530/585 nm is directly proportional to the D-amino acid concentration in the sample.
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 µL 1250 µM standard by mixing 125 µL 2 mM Standard and 75 µL dH2O.
For the sample plus standard well, add 5 µL 1250 µM standard and 20 µL sample.
For the sample and sample blank wells, add 5 µL dH2O and 20 µL sample.
Prepare enough Working Reagent (WR) for all assay wells by mixing, for each well, 85 µL Assay Buffer, 1 µL DAA Enzyme, 1 µL HRP Enzyme, 1 µL Dye Reagent. Prepare a WR without the DAA Enzyme for the sample blank well(s).
Add 80 µL of the WR to each well. Tap plate briefly to mix.
Incubate at room temperature for 60 min. Use a plate reader to read OD570nm.
The sample D-amino acid concentration is computed as follows:
where RSAMPLE, RBLANK, and RSTANDARD 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 4× lower than the sample volume; thus, the internal standard concentration should be divided by 4.
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