Mouse Dipeptidyl Peptidase 9 (DPP9) ELISA Kit

Principle of the Assay

The microtiter plate provided in this kit has been pre-coated with an antibody specific to DPP9. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to DPP9. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain DPP9, biotin-conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution, and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of DPP9 in the samples is then determined by comparing the O.D. of the samples to the standard curve.

For use with: Tissue homogenates and Other biological fluids.

For Research Use Only. Not for use in diagnostic procedures.

Target Information

Dipeptidyl peptidase that cleaves off N-terminal dipeptides from proteins having a Pro or Ala residue at position 2 (PubMed:24223149). Acts as a key inhibitor of caspase-1-dependent monocyte and macrophage pyroptosis in resting cells by preventing activation of NLRP1 and CARD8 (PubMed:27820798, PubMed:29396289). Sequesters the cleaved C-terminal part of NLRP1 and CARD8, which respectively constitute the active part of the NLRP1 and CARD8 inflammasomes, in a ternary complex, thereby preventing their oligomerization and activation (By similarity). The dipeptidyl peptidase activity is required to suppress NLRP1 and CARD8; however, neither NLRP1 nor CARD8 are bona fide substrates of DPP9, suggesting the existence of substrate(s) required for NLRP1 and CARD8 inhibition (By similarity).

GENE ID	224897
SWISS PROT	Q8BVG4
SYNONYMS	DPRP2; DPLP9; Dipeptidylpeptidase 9; Dipeptidyl peptidase IV-related protein 2; Dipeptidyl peptidase-like protein 9; Dipeptidyl peptidase IX

Materials Supplied

Kit Components	96 Wells Quantity/Size
Pre-coated, ready-to-use 96-well strip plate	1 plate
Plate sealer for 96 wells	2
Standard	2 tubes
Diluent buffer	1 bottle
Detection Reagent A	1 bottle
Detection Reagent B	1 bottle
TMB Substrate	1 tube
Stop Solution	1 tube
Wash Buffer (30X concentrate)	1 tube
Product data sheet	1 copy

Storage

Storage

The TMB Substrate, Wash Buffer (30X concentrate), and the Stop Solution should be stored at 4°C upon receipt, while the other items should be stored at -20°C.

Performance Characteristics

REPEATABILITY	Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level DPP9 were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle, and high-level DPP9 were tested on 3 different plates, with 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
SENSITIVITY	The minimum detectable dose was 25pg/mL.
ASSAY RANGE	78.1–5,000pg/mL
SPECIFICITY	This assay has high sensitivity and excellent specificity for the detection of DPP9. No significant cross-reactivity or interference between DPP9 and analogs was observed. Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between DPP9 and all analogs, therefore, cross reactivity may still exist.