EuroMAbNet: European Monoclonal Antibodies Network

mAbs Against HUMAN Protein

CD62P

STATUS:
CONTACT INFORMATION:	Immunology Unit, Faculty of Medicine and Medical Sciences, University of Barcelona
STATUS:	Validated
TYPE:	mouse anti human
CLONE NAME:	P.SEL KO 2.3
PROTEIN:	-
ANTIGEN USED:	300.19 transfected with human CD62P
FUSION PARTNER:	Ns1
ISOTYPE:	IgG1
SPECIES REACTIVITY:	-
PREPARATION AND STORAGE:	-
APP RECOMMENDED:	IP, Flow cytometry
APP NO TESTED:	IHQ-paraffin, IHQ-frozen, IF, WB

Description

P-selectin (CD62P) is an adhesion molecule expressed on the activated endothelium and activated platelets that is involved in the initial
attachment of leukocytes to inflamed vascular endothelium.
P-selectin is constitutively present in alfa-granules of platelets and Weibel-Palade bodies of endothelial cells, and is mobilized to the cell surface after activation by thrombogenic and inflammatory mediators.

References

Massaguer A, Engel P, Pérez-del-Pulgar S, Bosch J, Pizcueta P. Production and characterization of monoclonal antibodies against conserved epitopes of P-selectin (CD62P). Tissue Antigens. 2000, 56:117-128

Massaguer A, Perez-Del-Pulgar S, Engel P, Serratosa J, Bosch J, Pizcueta P. Concanavalin-A-induced liver injury is severely impaired in mice deficient in P-selectin. Leucocyte Typing VII.(ed. Mason D et al.), Oxford University Press, Oxford 2002, 342-343

Barrabés JA, Mirabet M, Agulló L, Figueras J, Pizcueta P, Garcia-Dorado D. Platelet deposition in remote cardiac regions after coronary occlusion. Eur J Clin Invest. 2007, 37:939-46

Mirabet M, Garcia-Dorado D, Inserte J, Barrabés JA, Lidón RM, Soriano B, Azevedo M, Padilla F, Agulló L, Ruiz-Meana M, Massaguer A, Pizcueta P, Soler-Soler J. Platelets activated by transient coronary occlusion exacerbate ischemia-reperfusion injury in rat hearts. Am J Physiol Heart Circ Physiol. 2002; 283:H1134-41

Massaguer A, Engel P, Tovar V, March S, Rigol M, Solanes N, Bosch J, Pizcueta P. Characterization of platelet and soluble-porcine P-selectin (CD62P). Vet Immunol Immunopathol. 2003, 96:169-81.

Applications

WB Techniques

WB Techniques	Clone	Dilution	Antibody concentration	+/- control	Expected MW	Observed Mw	Positivity in other species	Protocol
Western Blotting
Immunoprecipitation
	P.SEL KO 2.3	1 ug		/

Enlarge image

Immunoprecipitation of P-selectin from human platelets with P-sel.KO.2.2 and P-sel.KO.2.3 mAbs.

Human activated and surface biotinylated platelets were lysed and immunoprecipitated with P-sel.KO.2.2 and P-sel.KO.2.3 mAbs. A mAb against CD84 was used as positive control, and an irrelevant mAb as negative control. Samples were run on a SDS-PAGE gel under reducing conditions and blotted onto PVDF membrane.
Membrane was treated with HRP-avidin and the bands developed. Molecular weight markers (kDa) are indicated.

FLOW CYTOMETRY

FLOW CYTOMETRY	Clone	Dilution	+/- control	Type of fluorocrom	Protocol
	P.SEL KO 2.3	1 ug	/ untransfected 300.19 cells

Enlarge image

Specificity of the P-sel.KO.2.3 mAb for P.Selectin by flow cytometry

Representative histogram obtained after indirect immunofluorescence staining of untransfected 300.19
cells (dashed lines) and 300.19 cells transfected
with human P-selectin c-DNA (solid lines) with P-sel.KO.2.3. Fluorescence intensity is shown on a 4-decade log scale.