Human Cell Biology ELISA Kits 3
Human DPP4 (Dipeptidyl Peptidase IV) CLIA Kit (HUES00056)
- SKU:
- HUES00056
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 46.88pg/mL
- Range:
- 78.13-5000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- CD26, DPPIV, ADABP, ADCP2, TP103, Adenosine Deaminase Complexing Protein 2
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 78.13-5000 pg/mL |
| Sensitivity: | 46.88 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human DPP4 in samples. No significant cross-reactivity or interference between Human DPP4 and analogues was observed. |
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human DPP4. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human DPP4 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human DPP4, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human DPP4. The concentration of Human DPP4 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | DPP4: Cell surface glycoprotein receptor involved in the costimulatory signal essential for T-cell receptor (TCR)-mediated T-cell activation. Acts as a positive regulator of T-cell coactivation, by binding at least ADA, CAV1, IGF2R, and PTPRC. Its binding to CAV1 and CARD11 induces T-cell proliferation and NF- kappa-B activation in a T-cell receptor/CD3-dependent manner. Its interaction with ADA also regulates lymphocyte-epithelial cell adhesion. In association with FAP is involved in the pericellular proteolysis of the extracellular matrix (ECM), the migration and invasion of endothelial cells into the ECM. May be involved in the promotion of lymphatic endothelial cells adhesion, migration and tube formation. When overexpressed, enhanced cell proliferation, a process inhibited by GPC3. Acts also as a serine exopeptidase with a dipeptidyl peptidase activity that regulates various physiological processes by cleaving peptides in the circulation, including many chemokines, mitogenic growth factors, neuropeptides and peptide hormones. Removes N-terminal dipeptides sequentially from polypeptides having unsubstituted N-termini provided that the penultimate residue is proline. Belongs to the peptidase S9B family. DPPIV subfamily. |
| UniProt Protein Details: | Protein type:Cell surface; Membrane protein, integral; Cell adhesion; Protease; EC 3. 4. 14. 5 Chromosomal Location of Human Ortholog: 2q24. 3 Cellular Component: intercellular canaliculus; cell surface; focal adhesion; membrane; endocytic vesicle; lysosomal membrane; lamellipodium; apical plasma membrane; integral to membrane; plasma membrane; lipid raft Molecular Function:identical protein binding; protein binding; protein homodimerization activity; protease binding; dipeptidyl-peptidase activity; serine-type peptidase activity; serine-type endopeptidase activity; receptor binding Biological Process: regulation of cell-cell adhesion mediated by integrin; T cell activation; positive regulation of cell proliferation; T cell costimulation; response to hypoxia; endothelial cell migration; proteolysis |
| NCBI Summary: | The protein encoded by this gene is identical to adenosine deaminase complexing protein-2, and to the T-cell activation antigen CD26. It is an intrinsic membrane glycoprotein and a serine exopeptidase that cleaves X-proline dipeptides from the N-terminus of polypeptides. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P27487 |
| NCBI GenInfo Identifier: | 1352311 |
| NCBI Gene ID: | 1803 |
| NCBI Accession: | P27487. 2 |
| UniProt Secondary Accession: | P27487,Q53TN1, |
| UniProt Related Accession: | P27487 |
| Molecular Weight: | 88,279 Da |
| NCBI Full Name: | Dipeptidyl peptidase 4 |
| NCBI Synonym Full Names: | dipeptidyl-peptidase 4 |
| NCBI Official Symbol: | DPP4 |
| NCBI Official Synonym Symbols: | CD26; ADABP; ADCP2; DPPIV; TP103 |
| NCBI Protein Information: | dipeptidyl peptidase 4; ADCP-2; DPP IV; dipeptidylpeptidase 4; dipeptidyl peptidase IV; T-cell activation antigen CD26; adenosine deaminase complexing protein 2; dipeptidylpeptidase IV (CD26, adenosine deaminase complexing protein 2) |
| UniProt Protein Name: | Dipeptidyl peptidase 4 |
| UniProt Synonym Protein Names: | ADABP; Adenosine deaminase complexing protein 2; ADCP-2; Dipeptidyl peptidase IV; DPP IV; T-cell activation antigen CD26; TP103; CD_antigen: CD26Cleaved into the following 2 chains:Dipeptidyl peptidase 4 membrane formAlternative name(s):Dipeptidyl peptidase IV membrane form |
| Protein Family: | Dipeptidyl peptidase |
| UniProt Gene Name: | DPP4 |
| UniProt Entry Name: | DPP4_HUMAN |
As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (pg/mL) | RLU | Average | Corrected |
| 5000 | 56481 66031 | 61256 | 61233 |
| 2500 | 30929 37779 | 34354 | 34331 |
| 1250 | 18185 18153 | 18169 | 18146 |
| 625 | 8860 9924 | 9392 | 9369 |
| 312.5 | 5179 4487 | 4833 | 4810 |
| 156.25 | 2599 2423 | 2511 | 2488 |
| 78.13 | 1337 1341 | 1339 | 1316 |
| 0 | 22 24 | 23 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human DPP4 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human DPP4 were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 237.67 | 673.60 | 2467.34 | 224.47 | 722.14 | 2334.61 |
| Standard deviation | 30.28 | 50.39 | 201.09 | 25.77 | 56.98 | 157.35 |
| C V (%) | 12.74 | 7.48 | 8.15 | 11.48 | 7.89 | 6.74 |
Recovery
The recovery of Human DPP4 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 96-111 | 102 |
| EDTA plasma (n=5) | 95-109 | 102 |
| Cell culture media (n=5) | 88-101 | 94 |
Linearity
Samples were spiked with high concentrations of Human DPP4 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 92-106 | 101-116 | 89-100 |
| Average (%) | 97 | 107 | 95 | |
| 1:4 | Range (%) | 90-103 | 101-117 | 101-116 |
| Average (%) | 97 | 107 | 109 | |
| 1:8 | Range (%) | 91-106 | 89-104 | 99-114 |
| Average (%) | 97 | 95 | 106 | |
| 1:16 | Range (%) | 96-111 | 100-115 | 88-103 |
| Average (%) | 104 | 106 | 94 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro CLIA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- Determine the RLU value of each well immediately.
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