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.
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).
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Product Details
| Alternative Name: | CD26, Dipeptidyl Peptidase IV, DPP4 |
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| Sensitivity: | Can detect DPPIV activity in most or all biological samples. |
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| Applications: | Colorimetric detection, Fluorescent detection, HTS
Activity assay
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| Application Notes: | Correlation of DPPIV activity with disease states, or determination of the efficacy of DPPIV inhibitors administered in vivo. |
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| 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. |
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| Handling: | Keep frozen at all times. |
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| Shipping: | Dry Ice |
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| Long Term Storage: | -80°C |
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| Contents: | 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: -80°C; avoid freeze thaw cycles
pNA substrate (H-Gly-Pro-pNA)
(Prod. No. BML-P188-9091)
(55µl; 100mM in DMSO)
Storage: -80°C
pNA calibration standard (p-nitroaniline)
(Prod. No. BML-KI577)
(600μl; 80µM in 50mM Glycine, pH 8.7, 1mM EDTA)
Storage: -80°C
AMC substrate (H-Gly-Pro-AMC)
(Prod. No. BML-P189-9091)
(45µl; 50mM in DMSO),
Storage: -80°C
AMC calibration standard (7-amino-4-methylcoumarin)
(Prod. No. BML-KI578)
(600μl; 50µM in 50mM Glycine, pH 8.7, 1mM EDTA)
Storage: -80°C
Inhibitor P32/98
(Prod. No. BML-PI142-9090)
(20µl; 1mM in DMSO)
Storage: -80°C
Assay buffer
(Prod. No. BML-KI576)
(20ml; 50mM Glycine, pH 8.7, 1mM EDTA; liquid in screw-cap plastic bottle)
Storage: -80°C
½-volume clear microplate
(Prod. No. BML-KI101)
Storage: Ambient
½-volume NBS white microplate
(Prod. No. BML-KI571)
Storage: Ambient |
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| UniProt ID: | P27487 |
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| Regulatory Status: | RUO - Research Use Only |
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| Compatibility: | This product is compatible with the Absorbance 96 Plate Reader.
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Product Literature References
Abelmoschus esculentus subfractions ameliorate hepatic lipogenesis and lipid uptake via regulating dipeptidyl peptidase-4—With improving insulin resistance: C.H. Peng, et al.; PLoS One
17, e0265444 (2022),
Abstract;
Circulatory Adipokines and Incretins in Adolescent Idiopathic Scoliosis: A Pilot Study: E. Normand, et al.; Children
9, 1619 (2022),
Abstract;
Elevated dipeptidyl peptidase IV (DPP-IV) activity in plasma from patients with various lysosomal diseases: A. Lugowska, et al.; Diagnostics (Basel)
11, 320 (2021),
Abstract;
Full Text
A comprehensive evaluation of the impact of bovine milk containing different beta-casein profiles on gut health of ageing mice: B. Guantario, et al.; Nutrients
12, 2147 (2020),
Abstract;
Full Text
Abelmoschus esculentus subfractions attenuate Aβ and tau by regulating DPP-4 and insulin resistance signals: C. Huang, et al.; BMC Complement Med. Ther.
20, 370 (2020),
Abstract;
Full Text
Abelmoschus esculentus subfractions attenuate beta amyloid-induced neuron apoptosis by regulating DPP-4 with improving insulin resistance signals: C.N. Huang, et al.; PLoS One
14, e0217400 (2019),
Abstract;
Full Text
Linagliptin prevents atrial electrical and structural remodeling in a canine model of atrial fibrillation: T. Igarashi, et al.; Heart Vessels
33, 1258 (2018),
Abstract;
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
Saxagliptin Attenuates Albuminuria by Inhibiting Podocyte Epithelial- to-Mesenchymal Transition via SDF-1α in Diabetic Nephropathy: Y.P. Chang, et al.; Front. Pharmacol.
8, 780 (2017),
Application(s): Rat blood serum and renal tissue lysate,
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
In silico, in vitro and in vivo analyses of dipeptidyl peptidase IV inhibitory activity and the antidiabetic effect of sodium caseinate hydrolysate: C.H. Hsieh, et al.; Food Funct.
7, 1122 (2016),
Abstract;
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;
Improvement of glycemic control in streptozotocin-induced diabetic rats by Atlantic salmon skin gelatin hydrolysate as the dipeptidyl-peptidase IV inhibitor: C.H. Hsieh, et al.; Food Funct.
6, 1887 (2015),
Abstract;
Synthetic (+)‐antroquinonol exhibits dual actions against insulin resistance by triggering AMP kinase and inhibiting dipeptidyl peptidase IV activities: C.Y. Hsu, et al.; Br. J. Pharmacol.
172, 38 (2015),
Abstract;
Full Text
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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