P-selectin is a type-1 transmembrane protein that in humans is encoded by the SELP gene. P-selectin functions as a cell adhesion molecule (CAM) on the surfaces of activated endothelial cells, which line the inner surface of blood vessels, and activated platelets. In unactivated endothelial cells, it is stored in granules called Weibel-Palade bodies. In unactivated platelets P-selectin is stored in α-granules. Other names for P-selectin include CD62P, Granule Membrane Protein 140 (GMP-140), and Platelet Activation-Dependent Granule to External Membrane Protein (PADGEM). It was first identified in endothelial cells in 1989.
Background References
1. Hegazy S et al. CD62P (P-selectin) expression as a platelet activation marker in patients with liver cirrhosis with and without cholestasis. Clin Exp Hepatol. 2021 Jun
2. Takada YK et al. The C-type lectin domain of CD62P (P-selectin) functions as an integrin ligand. Life Sci Alliance. 2023 Apr
Western blot analysis of P-Selectin / CD62P on different lysates with Rabbit anti-P-Selectin / CD62P antibody (HA724039) at 1/2,000 dilution.
Lane 1: HeLa cell lysate (negative) (20 µg/Lane) Lane 2: HEL cell lysate (20 µg/Lane) Lane 3: Mouse placenta tissue lysate (40 µg/Lane) Lane 4: Mouse lung tissue lysate (40 µg/Lane) Lane 5: Rat placenta tissue lysate (40 µg/Lane) Lane 6: Rat lung tissue lysate (40 µg/Lane)
Predicted band size: 83 kDa Observed band size: 130-150 kDa
Exposure time: 1 minute; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA724039) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Western blot analysis of P-Selectin / CD62P on mouse platelets lysates with Rabbit anti-P-Selectin / CD62P antibody (HA724039) at 1/5,000 dilution.
Lysates/proteins at 30 µg/Lane.
Predicted band size: 83 kDa Observed band size: 150 kDa
Exposure time: 3 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA724039) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-P-Selectin / CD62P antibody (HA724039) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA724039) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-P-Selectin / CD62P antibody (HA724039) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA724039) at 1/337 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-P-Selectin / CD62P antibody (HA724039) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA724039) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Flow cytometric analysis of mouse platelets labeling P-Selectin / CD62P.
Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA724039, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Application: IHC-Fr
Species: Mouse
Site: spleen
Sample: Frozen section
Antibody concentration: 1/1,000
Antigen retrieval: Not required
P-Selectin / CD62P was immunoprecipitated from 0.2 mg HEL cell lysate with HA724039 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724039 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HEL cell lysate (input) Lane 2: HA724039 IP in HEL cell lysate Lane 3: Rabbit IgG instead of HA724039 in HEL cell lysate