P-Selectin ELISA Kit

Catalog No. ABIN411352

Product Details P-Selectin ELISA Kit

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

Binding Specificity: AA 42-709

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

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse P-Selectin

Brand: PicoKine™

Sample Type: Cell Culture Supernatant, Serum, Plasma (citrate)

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: W42-A709

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <5pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: NSO
Immunogen sequence: W42-A709

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Sequence similarities: Belongs to the selectin/LECAM family.

Tissue Specificity: Stored in the alpha-granules of platelets and Weibel-Palade bodies of endothelial cells. Upon cell activation by agonists, P-selectin is transported rapidly to the cell surface.

Plate: Pre-coated

Protocol: mouse P-Selectin ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for P-Selectin has been precoated onto 96-well plates. Standards(NSO, W42-A709) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for P-Selectin is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse P-Selectin amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 4000pg/mL, 2000pg/mL,1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL mouse P-Selectin standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of mouse cell culture supernates, serum or plasma(citrate) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse P-Selectin standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 325, Standard deviation: 14, CV(%): 4.3
• Sample 2: n=16, Mean(pg/ml): 874, Standard deviation: 43.7, CV(%): 5
• Sample 3: n=16, Mean(pg/ml): 1886, Standard deviation: 71.7, CV(%): 3.8,
• Sample 1: n=24, Mean(pg/ml): 417, Standard deviation: 27.11, CV(%): 6.5
• Sample 2: n=24, Mean(pg/ml): 1029, Standard deviation: 63.8, CV(%): 6.2
• Sample 3: n=24, Mean(pg/ml): 2020, Standard deviation: 94.94, CV(%): 4.7

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

References for P-Selectin ELISA Kit (ABIN411352)

Li, Ben, Yuan, Liu, Wu, Xu, Zhang: "Recruitment of HIV-1 target cells at topical mucosal sites: a sensitive and early marker for determining the safety of microbicide candidates." in: Emerging microbes & infections, Vol. 2, Issue 7, pp. e42, (2015) (PubMed).

Tun, Yasukawa, Yamada: "Involvement of nitric oxide with activation of Toll-like receptor 4 signaling in mice with dextran sodium sulfate-induced colitis." in: Free radical biology & medicine, Vol. 74, pp. 108-17, (2014) (PubMed).

Zhong, He, Yang, Bao, Zhang, Zhou, Chen, Li, Han, Yang, Sun, Cao, Li, Shi, Jiang, Zhang, Yan: "L-selectin and P-selectin are novel biomarkers of cervicovaginal inflammation for preclinical mucosal safety assessment of anti-HIV-1 microbicide." in: Antimicrobial agents and chemotherapy, Vol. 56, Issue 6, pp. 3121-32, (2012) (PubMed).

Yasukawa, Tokuda, Tun, Utsumi, Yamada: "The detrimental effect of nitric oxide on tissue is associated with inflammatory events in the vascular endothelium and neutrophils in mice with dextran sodium sulfate-induced colitis." in: Free radical research, Vol. 46, Issue 12, pp. 1427-36, (2012) (PubMed).

Target Details for P-Selectin

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

Alternative Name: SELP (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:

Protein Function: Ca(2+)-dependent receptor for myeloid cells that binds to carbohydrates on neutrophils and monocytes. Mediates the interaction of activated endothelial cells or platelets with leukocytes. The ligand recognized is sialyl-Lewis X. Mediates rapid rolling of leukocyte rolling over vascular surfaces during the initial steps in inflammation through interaction with PSGL1. .

Background: P-selectin, also called GMP-140, CD62, or selectin P, is a 140-kD adhesion molecule, expressed at the surface of activated cells, that mediates the interaction of activated endothelial cells or platelets with leukocytes. It is stored in secretory granules and expressed at the plasma membrane after cell activation. It is known to play an important role in atherosclerosis. The major ligand for P-selectin on leukocytes is P-selectin glycoprotein ligand-1(PSGL-1). The standard product used in this kit is recombinant mouse P-Selectin, excluding intercellular P-Selectin and transmembrane domain. It has 42-709 amino acids sequence with the molecular mass of 99.1KDa. As a result of glycosylation, the molecular mass of 190-200KDa is revealed by SDS-PAGE.

Synonyms: P-selectin,CD62 antigen-like family member P,Granule membrane protein 140,GMP-140,Leukocyte-endothelial cell adhesion molecule 3,LECAM3,Platelet activation dependent granule-external membrane protein,PADGEM,CD62P,Selp,Grmp,

Full Gene Name: P-selectin

Cellular Localisation: Membrane, Single-pass type I membrane protein.

Gene ID: 20344

UniProt: Q01102