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Human CD26 / DPP4 ELISA Kit

SKU:
HUFI00003
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P27487
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
ELISA Type:
Sandwich
Synonyms:
CD26, DPP4, CD26, ADABP, ADCP-2, ADCP2DPP IV, Adenosine deaminase complexing protein 2TP103, CD26 antigen, CD26T-cell activation antigen CD26, dipeptidyl peptidase 4, Dipeptidyl peptidase IV, dipeptidylpeptidase 4, dipeptidyl-peptidase 4, dipeptidylp
Reactivity:
Human
Research Area:
Cell Biology
$719
Frequently bought together:

Description

Human CD26/DPP4 ELISA Kit

The Human CD26 (DPP4) ELISA Kit is a highly sensitive and specific assay designed for the accurate quantification of CD26 levels in human serum, plasma, and cell culture supernatants. CD26, also known as Dipeptidyl peptidase-4 (DPP4), is a cell surface glycoprotein that plays a critical role in immune regulation, T-cell activation, and inflammation.This ELISA kit offers reliable and reproducible results, making it an essential tool for researchers studying autoimmune diseases, cancer, and metabolic disorders where CD26 is implicated.

By accurately measuring CD26 levels, researchers can gain valuable insights into disease pathology, progression, and potential therapeutic targets. Take your research to the next level with the Human CD26 (DPP4) ELISA Kit from Assay Genie, the trusted choice for high-quality, innovative research tools. Order yours today and unlock the power of precision in your experiments.

Product Name:Human CD26 / DPP4 ELISA Kit
Product Code:HUFI00003
Size:96 Assays
Alias:CD26, DPP4, CD26, ADABP, ADCP-2, ADCP2DPP IV, Adenosine deaminase complexing protein 2TP103, CD26 antigen, CD26T-cell activation antigen CD26, dipeptidyl peptidase 4, Dipeptidyl peptidase IV, dipeptidylpeptidase 4, dipeptidyl-peptidase 4, dipeptidylpeptidase IV, CD26, adenosine deaminase complexing protein 2, DPPIV
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human CD26 concentrations in serum plasma and other biological fluids.
Sensitivity:0.188ng/ml
Range:0.313-20ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human CD26 and the recovery rates were calculated by comparing the measured value to the expected amount of Human CD26 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)88-10396
EDTA plasma(n=5)87-9993
UFH plasma(n=5)87-104100
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human CD26 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-104%88-105%86-104%
EDTA plasma(n=5)85-95%82-96%86-101%
UFH plasma(n=5)83-98%87-98%81-95%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotP27487
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

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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