P-Selectin ELISA Kit

Catalog No. ABIN1112781

Product Details P-Selectin ELISA Kit

Target: P-Selectin (SELP) (Selectin P (Granule Membrane Protein 140kDa, Antigen CD62) (SELP))

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5-4000 pg/mL

Minimum Detection Limit: 62.5 pg/mL

Application: ELISA

Analytical Method: Quantitative

Sensitivity: < 5 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Mouse P-Selectin antibody 2. Lyophilized P-Selectin standards: 2 tubes (20ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Mouse P-Selectin antibody (Concentrated): 130 µl.

Material not included: 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Application Details

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- P-Selectin polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- P-Selectin polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the P-Selectin amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of P-Selectin can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Tissue lysate or body fluids, cell culture supernate: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 2000 × g for 20 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with EDTA as the anticoagulant. Centrifuge for 20 min at 2000 x g within 30 min of collection. Analyze immediately or aliquot and store frozen at -20 °C. Heparin and citrate can not be used as anticoagulant here. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Target Details for P-Selectin

Target: P-Selectin (SELP) (Selectin P (Granule Membrane Protein 140kDa, Antigen CD62) (SELP))

Alternative Name: P-Selectin (SELP Products)

Synonyms: PSELECT ELISA Kit, CD62 ELISA Kit, CD62P ELISA Kit, GMP140 ELISA Kit, GRMP ELISA Kit, LECAM3 ELISA Kit, PADGEM ELISA Kit, PSEL ELISA Kit, GMP-140 ELISA Kit, Grmp ELISA Kit, selectin P ELISA Kit, selectin, platelet ELISA Kit, Selp ELISA Kit, SELP ELISA Kit

Background: P-selectin, also called GMP-140, CD62, or selectin P, is a 140-kD adhesion molecule that in humans is encoded by the SELP gene. It is located on chromosome 1q21-q24, spans>50 kb and contains 17 exons in human. It is constitutively expressed on megakaryocytes (the precursor of platelets) and endothelial cells. P-selectin plays an essential role in the initial recruitment of leukocytes (white blood cells) to the site of injury during inflammation. Ligands for P-selectin on eosinophils and neutrophils are similar sialylated, protease-sensitive, endo-beta-galactosidase-resistant structures, clearly different than those reported for E-selectin, and suggest disparate roles for P-selectin and E-selectin during recruitment during inflammatory responses.