EnzyChrom™ D-Amino Acid Assay Kit

Application

• For quantitative determination of D-Amino Acids in biological and other samples.

Key Features

• 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.

Method

• OD570nm or FL530/585nm

Samples

• Tissue, milk, and other biological samples

Species

• All

Procedure

• 60 min

Size

• 100 tests

Detection Limit

• 0.86 µM (colorimetric assay), 0.18 µM (fluorimetric assay)

Shelf Life

• 6 months

More Details

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 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 4× lower than the sample volume; thus, the internal standard concentration should be divided by 4.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.