GMP-140 (P-selectin) EIA Kit (GWB-CB73D4)





Product Name: GMP-140 (P-selectin) EIA Kit

# of Samples: 1X96 Assays

Intended Use: The GMP-140 EIA Kit is an in vitro enzyme immunoassay (EIA) kit for quantitative determination of human GMP-140 in plasma, cultured cell extracts, cell culture supernatants, and other biological fluids.
This kit is for research use only. It is not for use in diagnostic or therapeutic procedures.

Introduction: GMP-140, also known as CD62 or PADGEM (6), is a 140 kDa integral membrane glycoprotein functionally categorized as a member of the LEC-CAM family (lectinepidermal growth factor-complement binding cell adhesion molecules), or the selectin family. In non-activated conditions, GMP-140 is present in the alpha-granules of platelets or in the Weibel Palade bodies of endothelial cells (1-5). However, when activated by stimulators such as thrombin or by inflammatory or hemostatic mediators during intravascular inflammation, GMP-140 is rapidly secreted to the plasma membrane to promote adhesion of cells to vascular endothelia or to neutrophils and monocytes at the site of tissue injury (10-12). Previous studies suggested that GMP-140 is indeed a useful marker of platelet and endothelium activation in experimental inflammation models, and potentially in clinical situations (5, 7-9). GMP-140 is also known to interact with leukocytes by a Ca2+ dependent lectin-like mechanism (13, 14). GMP-140 has recently been cloned from endothilia and is found to be highly homologous to two other LEC-CAM adhesion molecules, ELAM-1 and the Mel-14 antigen, both of which are involved in various aspects of leukocyte adhesion (3, 15). Analysis of the endothelium cDNA suggests that there are three variants of GMP-140 resulting from alternative splicing (16): two transmembrane forms with different numbers of compliment-binding consensus repeats, and one putative cytoplasmic “soluble form” that lacks the hydrophobic transmembrane domain (20). Recently it was shown that GMP-140 molecules extracted by Triton X-100 from platelet membranes polymerize into soluble tetramers when returned to detergent-free conditions, and that this tetramer strongly inhibits the adhesion of activated neutrophils to endothelia. This is evidence that GMP-140-mediated adhesion works via a saturable GMP-140 receptor on the surface of myeloid cells (18). This also suggests that if free GMP-140 molecules are at all present in the circulation, they may serve to maintain the nonadhesiveness of neutrophils and prevent the development of inflammatory responses (17).
The present Enzyme Immunoassay Method (EIA) was developed for qualitative and quantitative detection of GMP-140 by using two different monoclonal antibodies (19,21). Advantages of this EIA system are speed, simplicity, and high specificity.

Principle of the test: The GMP-140 EIA Kit is a solid phase EIA based on a sandwich method that utilizes two mouse monoclonal anti GMP-140 antibodies to detect GMP-140 by two-step procedure. One of the mouse monoclonal anti GMP-140 has been pre-coated onto a microtiter-plate and blocked against non-specific binding. Samples and standards are incubated in microtiter-plate wells. The second step is to wash the plate and add a second anti GMP-140 labelled with peroxidase (POD). During the incubation, GMP-140 is bound to anti GMP-140 (solid phase) on one side and tagged on the other by POD-anti GMP-140. The reaction between POD and substrate (H2O2 and tetramethylbenzidine) results in colour development with intensities proportional to the amount of GMP-140 present in samples and standards. The amount of GMP- 140 can be quantitated by measuring the absorbance using an EIA plate reader. Accurate sample concentrations of GMP-140 can be determined by comparing their specific absorbances with those obtained for the standards plotted on a standard curve.

Storage and Stability: This kit is shipped at 2 - 8C and should be stored at 2 - 8C if not used. Under this condition, the kit is stable until the expiration date on label.

Sensitivity and Specificity: This kit specifically measures GMP-140 with no detectable cross reaction with platelet Gp Ib, platelet Gp IIb, platelet Gp IIIa, von Willebrand factor, and platelet thrombospondin. This kit cannot be used to measure mouse GMP- 140. The application of this kit for quantitating GMP-140 from other sources has not been tested.

Product Name: GMP-140 (P-selectin) EIA Kit

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.

Subunit: Interacts with SNX17. Interacts with PSGL1/SEPL and mediateS neutrophil adhesion and leukocyte rolling. This interaction requires the sialyl-Lewis X epitope and specific tyrosine sulfation on PSGL1.

Subcellular Location: Membrane; Single-pass type I membrane protein.

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.

Disease: Defects in SELP may be a cause of susceptibility to ischemic stroke [MIM:601367]; also known as cerebrovascular accident or cerebral infarction. A stroke is an acute neurologic event leading to death of neural tissue of the brain and resulting in loss of motor, sensory and/or cognitive function. Ischemic strokes, resulting from vascular occlusion, is considered to be a highly complex disease consisting of a group of heterogeneous disorders with multiple genetic and environmental risk factors.

Similarity: Belongs to the selectin/LECAM family.

Similarity: Contains 1 C-type lectin domain.

Similarity: Contains 1 EGF-like domain.

Similarity: Contains 9 Sushi (CCP/SCR) domains.

Additional Information

Name GMP-140 (P-selectin) EIA Kit (GWB-CB73D4)
Related Product Names GMP-140 (P-selectin) EIA ELISA Kit; Granule membrane protein 140; GMP-140; PADGEM; Leukocyte-endothelial cell adhesion molecule 3; LECAM3; CD62 antigen-like family member P; CD62P antigen GMRP; GRMPGMP-140 (P-selectin) EIA KitSELP
Swissprot ID P16109
NCBI Acc Number NP_002996.2
Molecular Weight 90834
Datasheets/Manuals Printable datasheet for GWB-CB73D4
Intended Use Research Use Only