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DPPIV/CD26 Assay kit for biological samples

 
BML-AK498-0001 96 wells 425.00 USD
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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.
BML-AK498 activity1
BML-AK498 activity2A
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.
BML-AK498 activity2B
Twenty microliters of unconcentrated urine or saliva were assayed at 25°C (30 minute read) with GP-AMC (200µM).
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BML-AK498 activity1 BML-AK498 activity2A BML-AK498 activity2B

Product Specification

Alternative Name:CD26, Dipeptidyl Peptidase IV, DPP4
 
Sensitivity:Can detect DPPIV activity in most or all biological samples.
 
Applications:Colorimetric detection, Fluorescent detection, HTS
Activity assay
 
Application Notes:Correlation of DPPIV activity with disease states, or determination of the efficacy of DPPIV inhibitors administered in vivo.
 
Use/Stability:Substrates and inhibitor are in DMSO and are stable for up to 6 months. DPPIV enzyme activity decreases with time, even at -80°C.
 
Handling:Keep frozen at all times.
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Kit/Set Contains:DPPIV enzyme (Human, recombinant) (Prod. No. BML-SE434-9091) (5 mU; One U=1μmole/min at 37°C, 100µM H-Gly-Pro-pNA. Purity ≥95% (SDS-Page)), Storage: -70°C; avoid freeze thaw cycles
pNA substrate (H-Gly-Pro-pNA) (Prod. No. BML-P188-9091) (55µl; 100mM in DMSO)Storage: -70°C
pNA calibration standard (p-nitroaniline) (Prod. No. BML-KI577) (600μl; 80µM in 50mM Glycine, pH 8.7, 1mM EDTA)Storage: -70°C
AMC substrate (H-Gly-Pro-AMC) (Prod. No. BML-P189-9091) (45µl; 50mM in DMSO), Storage: -70°C
AMC calibration standard (7-amino-4-methylcoumarin) (Prod. No. BML-KI578)(600μl; 50µM in 50mM Glycine, pH 8.7, 1mM EDTA), Storage: -70°C
Inhibitor P32/98 (Prod. No. BML-PI142-9090) (20µl; 1mM in DMSO), Storage: -70°C
Assay buffer (Prod. No. BML-KI576) (20ml; 50mM Glycine, pH 8.7, 1mM EDTA; liquid in screw-cap plastic bottle), Storage: -20 or -70°C
½-volume clear microplate (Prod. No. BML-KI101), Storage: Room temperature
½-volume NBS white microplate (Prod. No. BML-KI571), Storage: Room temperature
 
UniProt ID:P27487
 

Product Literature References

Encapsulation of 16-hydroxycleroda-3,13-dine-16,15-olide in mesoporous silica nanoparticles as a natural dipeptidyl peptidase-4 inhibitor potentiated hypoglycemia in diabetic mice: P.K. Huang, et al.; Nanomaterials 7, E112 (2017), Application(s): DPPIV activity in Caco-2 cells, Abstract; Full Text
Inhibition of dipeptidyl peptidase-4 ameliorates cardiac ischemia and systolic dysfunction by up-regulating the FGF-2/EGR-1 pathway: M. Suda , et al.; PLoS One 12, e0182422 (2017), Application(s): Plasma samples from mice, Abstract; Full Text
Hypoxia alters the expression of dipeptidyl peptidase 4 and induces developmental remodeling of human preadipocytes: H.H. Chowdhury, et al.; J. Diabetes. Res. 2016, 7481470 (2016), Application(s): Soluble DPPIV activity in culture supernatant, Abstract; Full Text
Monitoring of dipeptidyl peptidase-IV (DPP-IV) activity in patients with mucopolysaccharidoses types I and II on enzyme replacement therapy - Results of a pilot study: K. Hetmariczyk, et al.; Clin. Biochem. 49, 458 (2016), Application(s): Measured plasma DPP-IV, Abstract;
Effect of dipeptidyl peptidase-4 inhibition on circadian blood pressure during the development of salt-dependent hypertension in rats: A. Sufiun, et al.; Hypertens. Res. 38, 237 (2015), Application(s): Assay, Abstract;
Dietary A1 β-casein affects gastrointestinal transit time, dipeptidyl peptidase-4 activity, and inflammatory status relative to A2 β-casein in Wistar rats: M.P. Barnett, et al.; Int. J. Food Sci. Nutr. 65, 720 (2014), Abstract;
Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC: V.S. Raj, et al.; Nature 495, 251 (2013), Abstract;
Dipeptidyl peptidase-4 inhibitor anagliptin facilitates restoration of dextran sulfate sodium-induced colitis: S. Mimura, et al.; Scand. J. Gastroenterol. 48, 1152 (2013), Abstract;

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