710281

antibody from Invitrogen Antibodies

Targeting: SELP CD62, CD62P, GMP140, GRMP, PADGEM, PSEL

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

710281

Provider

Invitrogen Antibodies

Product name

P-Selectin Recombinant Polyclonal Antibody (3HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with non-human primate, mouse and rat based on sequence homology.

Antibody clone number

3HCLC

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of P-selectin in Jurkat whole cell lysate using a P-selectin Recombinant Rabbit Polyclonal Antibody (Product # 710281) at a dilution of 1 µg/mL. Samples were detected using chemiluminescence (ECL). Results show a band at ~140kDa.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of P-selectin/CD62P was performed by loading 30 µg of K562, HEK-293, U-2 OS, A549, MCF-7, HeLa, Jurkat, A431, and PC3 cell lysates using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Proteins were transferred to a PVDF membrane and blocked with 5% skim milk for 1 hour at room temperature. P-selectin/CD62P was detected at ~140 kDa using P-selectin/CD62P Recombinant Rabbit Polyclonal Antibody (Product # 710281) at a 1:1000 dilution in 2.5% skim milk at 4°C overnight on a rocking platform. Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody (Product # G-21234) at a 1:5000 dilution and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of P-selectin/CD62P was performed on 70% confluent log phase U-2 OS cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0. 25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with P-selectin/CD62P Recombinant Rabbit Polyclonal Antibody (Product # 710281) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Goat anti-Rabbit IgG secondary antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization. The images were captured using a Nikon microscope at 20X magnification.