| DSP Rapid Kit DSP: Diarrhoeic Shellfish Poisoning (A colorimetric phosphatase inhibition assay) |
|
 |
About DSP
Assay Principle
The assay is based on the inhibition of the protein phosphatase (PP2A) by DSP toxins (OA and DTXs). PP2A can hydrolyze a colorless artificial substrate, p-nitrophenyl phosphate (p-NPP), and produces the yellow color of p-nitrophenol (p-NP) in the alkaline solution. The intensity of the color is proportional to the enzyme activity and the absorbance is measured at 405 nm. The concentration of DSP toxins in the sample is calculated from the standard curve produced using known concentrations of OA.
>> top
Summary of Assay
 |
96 Well plate | |
|---|---|---|
 |
Add 50 μl of Sample and OA Reference Solution |
|
|
Add 100 μl of PP2A Working Solution |
||
|
Add 100 μl of Substrate Working Solution |
||
| Mix for 1 min by a microplate mixer | ||
 |
||
| Incubate for 30 min at 36℃ | ||
|
||
| Measure absorbance at 405 nm against 490 or 492nm as reference. | ||
>> top
Advantages
- Simple step and Very sensitive assay
lowest quantifiable limit 0.1 μg/g DG*
*DG : digestive gland - High Accuracy and Reproducibility
>> top
Kit Components
- Okadaic Acid (OA) Reference Solutions (0, 0.5, 1.0, 1.5, 2.0, 2.5, 5.0 and 10.0 ng/ml)
- Sample Buffer
- 1.25 N NaOH Solution
- 1.25 N HCl Solution
- PP2A Buffer A
- PP2A Buffer B
- PP2A Stock Solution (0.5 ml tube)
- Substrate Buffer
- Substrate Tablet (p-NPP )
- 96 Well plate
- Adhesive Film
>> top
Usage (Animation)
>> top
Contact us
Tropical Technology Center Ltd.
5-1
Suzaki Uruma Okinawa, 904-2234, Japan
Tel: +81-98-982-1100, Fax: +81-98-982-1101
contact
us , URL : http://www.ttc.co.jp/dsp/
>> top