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| Product Name | Dipeptidyl Peptidase IV (DPP-4) Activity Fluorometric Assay Kit |
| Catalog No. | ETR-HMM-0086 |
| Description | Fluorometric assay for DPP-4 (CD26) activity using Gly-Pro-AMC as the specific fluorogenic substrate. DPP-4 is a serine exopeptidase that cleaves X-Pro or X-Ala dipeptides from the N-terminus of polypeptides. This enzyme is a key therapeutic target in type 2 diabetes (DPP-4 inhibitors: sitagliptin, vildagliptin) and is also involved in immune regulation and tumor biology. |
| Intended Use | DPP-4 enzyme activity measurement in serum/plasma, cell lysates, and purified enzyme; DPP-4 inhibitor screening and IC50 determination; diabetes drug discovery; clinical DPP-4 activity biomarker measurement; recombinant DPP-4 quality control. |
| Principle / Technology | DPP-4 cleaves Gly-Pro-AMC → Gly-Pro + AMC (fluorescent, Ex 360/Em 460 nm); inhibitors (sitagliptin, vildagliptin, saxagliptin) reduce AMC release; fluorescence proportional to enzyme activity |
| Detection Method | Fluorometric; Ex 350-360 nm, Em 440-460 nm; kinetic or endpoint; 96-well or 384-well format |
| Sample Type | Human/animal serum or plasma (EDTA or heparin; citrate inhibits DPP-4), cell lysates, purified recombinant DPP-4; 5-25 µL serum per assay |
| Sensitivity / LOD | LOD 0.1 mU/mL DPP-4; linear range 0.1-100 mU/mL in serum; detects DPP-4 activity in diluted (1:10) human serum |
| Specificity | Highly specific for DPP-4; DPP-8 and DPP-9 also cleave Gly-Pro-AMC but require different pH optima and inhibitor profiles; include sitagliptin control (DPP-4 selective) vs. DPP-8/9 inhibitor (1G244) for selectivity confirmation |
| Reaction Conditions / Protocol | Mix sample with assay buffer ± inhibitor (10 min pre-incubation); add Gly-Pro-AMC substrate (50 µM final); read fluorescence kinetically for 20-30 min at 37 °C; calculate DPP-4 activity from AMC standard curve |
| Components / Formulation | Gly-Pro-AMC substrate (10 mM in DMSO), AMC fluorescence standard (1 mM in DMSO), DPP-4 assay buffer (Tris-HCl 100 mM, NaCl 100 mM, pH 8.0), sitagliptin (DPP-4 inhibitor positive control, 10 mM in DMSO), purified human DPP-4 (positive control, 10 mU), black 96-well plate (optional) |
| Storage Conditions | -20 °C (substrate, AMC, DPP-4 enzyme, sitagliptin); assay buffer at 4 °C; protect from light |
| Shelf Life | 12 months at -20 °C |
| Package Specifications | 100 assays, 500 assays (96-well); HTS format available |
| Product Form | Lyophilized DPP-4 enzyme; liquid substrate and inhibitors in DMSO |
| Key Features | DPP-4-specific Gly-Pro-AMC substrate; includes sitagliptin inhibitor for specificity validation; AMC standard for quantification; DPP-4 positive control; suitable for serum and plasma clinical samples; DMSO-tolerant up to 2%; compatible with HTS formats |
| Purity | Gly-Pro-AMC >98% HPLC; AMC standard >99%; DPP-4 specific activity >30 U/mg; sitagliptin >98% |
| Concentration | As specified per kit; calibrator and substrate concentrations traceable to reference |
| Activity / Unit Definition | One unit defined as the amount of enzyme that catalyzes conversion of 1 µmol substrate per minute under specified conditions |
| Molecular Weight | Varies by target enzyme as documented |
| Source / Origin | Recombinant or purified from natural sources; synthetic chromogenic/fluorogenic substrates |
| pH Range / Optimal pH | pH 6.0–9.0 depending on specific enzyme assay |
| Shipping Conditions | Cold pack -20 °C; protect fluorogenic materials from light |
| Expiration Date / Stability | 12 months at -20 °C; reconstituted DPP-4 1 week at 4 °C; substrate DMSO stock 6 months at -20 °C |
| Regulatory / Compliance | For laboratory and research use only; RUO; sitagliptin is a prescription drug — for laboratory use only; ISO 9001 |
| Compatibility | Compatible with standard microplate readers (absorbance, fluorescence, luminescence modes) |
| Recommended Buffer System | Tris or phosphate buffer optimized per enzyme assay |
| Application Notes / Precautions | Include positive control and inhibitor control in each assay; read within linear range; protect light-sensitive substrates |
| Batch-to-Batch Consistency | Enzyme activity and standard curve linearity within ±15% of reference lot |
For research use only, not for clinical use.
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