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Rat CD26 / DPPIV ELISA Kit

SKU:
RTFI00738
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P14740
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
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, dipeptidylpeptidase IV, CD26, adenosine d
Reactivity:
Rat
Research Area:
Cell Biology
€649
Frequently bought together:

Description

system_update_altDatasheetsystem_update_altMSDS
Product Name:Rat DPP? (Dipeptidyl Peptldase ?) ELISA Kit
Product Code:RTFI00738
Size:96 Assays
Target:Rat DPP?
Alias: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, dipeptidylpeptidase IV, CD26, adenosine deaminase complexing protein 2, DPPIV, DPP?
Reactivity:Rat
Detection Method:Sandwich ELISA, Double Antibody
Sensitivity:0.094ng/ml
Range:0.156-10ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Rat DPP? and the recovery rates were calculated by comparing the measured value to the expected amount of Rat DPP? in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)86-10495
EDTA plasma(n=5)95-10398
UFH plasma(n=5)93-10598
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat DPP? and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)93-103%85-104%85-102%
EDTA plasma(n=5)87-97%84-97%83-100%
UFH plasma(n=5)80-99%81-99%84-97%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:P14740
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:Motility/polarity/chemotaxis; Endoplasmic reticulum; Cell surface; Cell adhesion; Membrane protein, integral; Protease; Cell cycle regulation; Vesicle; EC 3.4.14.5

Cellular Component: Golgi apparatus; intercellular canaliculus; cell surface; focal adhesion; endoplasmic reticulum; lysosomal membrane; integral to membrane; extracellular region; lipid raft; membrane; lamellipodium; endocytic vesicle; apical plasma membrane; plasma membrane; cell junction

Molecular Function:collagen binding; identical protein binding; protein homodimerization activity; protease binding; serine-type peptidase activity; dipeptidyl-peptidase activity; serine-type endopeptidase activity; peptide binding; receptor binding

Biological Process: regulation of cell-cell adhesion mediated by integrin; behavioral fear response; T cell activation; establishment of localization; regulation of T cell mediated immunity; positive regulation of cell proliferation; T cell costimulation; response to hypoxia; endothelial cell migration; cell adhesion; proteolysis

NCBI Summary:catalyzes the degradation of glucagon-like peptide-1 (Glp-1); involved in proteolysis and peptidolysis [RGD, Feb 2006]
UniProt Code:P14740
NCBI GenInfo Identifier:56405289
NCBI Gene ID:25253
NCBI Accession:P14740.2
UniProt Related Accession:P14740
Molecular Weight:88,089 Da
NCBI Full Name:Dipeptidyl peptidase 4
NCBI Synonym Full Names:dipeptidylpeptidase 4
NCBI Official Symbol:Dpp4  
NCBI Official Synonym Symbols:CD26; DPPIV  
NCBI Protein Information:dipeptidyl peptidase 4; DPP IV; GP110 glycoprotein; dipeptidyl peptidase IV; T-cell activation antigen CD26; bile canaliculus domain-specific membrane glycoprotein
UniProt Protein Name:Dipeptidyl peptidase 4
UniProt Synonym Protein Names:Bile canaliculus domain-specific membrane glycoprotein; Dipeptidyl peptidase IV; DPP IV; GP110 glycoprotein; T-cell activation antigen CD26; CD_antigen: CD26Cleaved into the following 3 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_RAT
StepProcedure
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 Type Protocol
Serum:

If 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 clotovernight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Removeserum and assay promptly or aliquot and store the samples at-80°C. Avoid multiple freeze-thaw cycles.

Plasma: Collect plasma using EDTA or heparin as an anti-coagulant. 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 Fluid: Collect 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 Supernatant: Collect 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 Lysates: Solubilize 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 Homogenates: The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenizein 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 Lysates: Rinse 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 Milk: Collect 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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