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Because DPPIV/CD26 protein levels may not accurately reflect the levels of active DPPIV enzyme, it is useful to measure DPPIV activity rather than performing DPPIV immunoassay. The DPPIV/CD26 Assay kit for biological samples is a complete assay system designed to measure DPPIV activity in biological fluids such as plasma, serum, urine, and saliva. Uses for the kit include correlation of DPPIV activity with disease states or determination of the efficacy of DPPIV inhibitors administered in vivo. The kit can easily be used for other biological fluids such as tissue, live cells and cell extracts, and exudates. Enough reagents are provided to perform at least 96 assays. The kit contains both a chromogenic substrate (H-Gly-Pro-pNA) and a fluorogenic substrate (H-Gly-Pro-AMC). Cleavage of the p-nitroaniline (pNA) from the chromogenic substrate increases absorbance at 405 nm. The fluorimetric assay is based on the cleavage of 7-amino-4-methylcoumarin (AMC) moiety from the C-terminus of the peptide substrate, which increases its fluorescence intensity at 460 nm. A specific DPPIV inhibitor, P32/98, is included to uncover any activity not contributed by DPPIV. Also included is DPPIV enzyme to use as a positive control.
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Twenty microliters of biological sample were assayed at 25°C (30 minute read at 380/460nm) with GP-AMC (200 µM final). Negative control contained only substrate in assay buffer. Possible GP-AMC substrate depletion by undiluted plasma and serum became apparent after approximately 13 minutes. Unconcentrated urine and saliva activity levels were 40 and 80 times lower than plasma or serum, but readily detectable.
Twenty microliters of unconcentrated urine or saliva were assayed at 25°C (30 minute read) with GP-AMC (200µM).
DPPIV/CD26 Assay kit for biological samples activity1
| Regulatory Status |
RUO – Research Use Only |
|---|
Related Products
| Application | Activity assay, Colorimetric detection, Fluorescent detection, HTS |
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| Alternative Name | Dipeptidyl Peptidase IV, DPP4, CD26 |
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| Purity | ≥95% (SDS-PAGE) |
| Source | Produced in Sf9 insect cells using baculovirus expression system. Active recombinant human DPPIV (aa 29-766). This represents a naturally-occurring cleaved (soluble) form of DPPIV. |