14-0627-82

antibody from Invitrogen Antibodies

Targeting: SELE CD62E, ELAM, ELAM1, ESEL

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Recommended by provider)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

14-0627-82

Provider

Invitrogen Antibodies

Product name

CD62E (E-selectin) Monoclonal Antibody (P2H3), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The P2H3 monoclonal antibody reacts with human CD62E, a 97-115 kDa member of the selectin family. CD62E, also known as E-selectin or endothelial-leukocyte adhesion molecule-1 (ELAM-1) is an adhesion molecule expressed by endothelial cells upon stimulation with cytokines including TNFalpha and IL-1beta. Induced expression of CD62E during inflammatory conditions is thought to mediate leukocyte rolling including the initial interaction of neutrophils with endothelium. The P2H3 monoclonal antibody also inhibits cellular adhesion to cytokine-activated endothelial cells. Applications Reported: This P2H3 antibody has been reported for use in flow cytometric analysis, immunoprecipitation, and immunohistochemical staining. Applications Tested: This P2H3 antibody has been tested by flow cytometric analysis of TNF alpha-actiavted Human Umbilical Vein Endothelial Cells (HUVEC). This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

P2H3

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

4° C

References

Submitted references

Surgical and intravital microscopy protocol to image Trypanosoma brucei-host interactions in live rodent models.

De Niz M, Figueiredo LM
STAR protocols 2022 Jun 17;3(2):101450

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Organotypic endothelial adhesion molecules are key for Trypanosoma brucei tropism and virulence.

De Niz M, Brás D, Ouarné M, Pedro M, Nascimento AM, Henao Misikova L, Franco CA, Figueiredo LM
Cell reports 2021 Sep 21;36(12):109741

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A Norrin/Wnt surrogate antibody stimulates endothelial cell barrier function and rescues retinopathy.

Chidiac R, Abedin M, Macleod G, Yang A, Thibeault PE, Blazer LL, Adams JJ, Zhang L, Roehrich H, Jo HN, Seshagiri S, Sidhu SS, Junge HJ, Angers S
EMBO molecular medicine 2021 Jul 7;13(7):e13977

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Colonization of dermal arterioles by Neisseria meningitidis provides a safe haven from neutrophils.

Manriquez V, Nivoit P, Urbina T, Echenique-Rivera H, Melican K, Fernandez-Gerlinger MP, Flamant P, Schmitt T, Bruneval P, Obino D, Duménil G
Nature communications 2021 Jul 27;12(1):4547

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Using CRISPR-Cas9 to quantify the contributions of O-glycans, N-glycans and Glycosphingolipids to human leukocyte-endothelium adhesion.

Stolfa G, Mondal N, Zhu Y, Yu X, Buffone A Jr, Neelamegham S
Scientific reports 2016 Jul 26;6:30392

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Effect of nicotine and porphyromonas gingivalis lipopolysaccharide on endothelial cells in vitro.

An N, Andrukhov O, Tang Y, Falkensammer F, Bantleon HP, Ouyang X, Rausch-Fan X
PloS one 2014;9(5):e96942

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Endothelial leukocyte adhesion molecule 1: an inducible receptor for neutrophils related to complement regulatory proteins and lectins.

Bevilacqua MP, Stengelin S, Gimbrone MA Jr, Seed B
Science (New York, N.Y.) 1989 Mar 3;243(4895):1160-5

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-CD62E (E-selectin) Monoclonal Antibody (Product # 14-0627-82) and 67, 115 kDa bands corresponding to CD62E were observed across the tissues tested. Tissue extracts (30 µg lysate) of Mouse Liver (Lane 1), Mouse Spleen (Lane 2) and Mouse White Adipose (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Staining of TNF alpha-stimulated Human Umbilical Vein Endothelial Cells (HUVEC) with 0.5 µg of Mouse IgG1 kappa Isotype Control Purified (Product # 14-4714-82) (open histogram) or 0.5 µg of Anti-Human CD62E (E- Selectin) Purified (filled histogram) followed by F (ab')2 Anti-Mouse IgG PE (Product # 12-4012). Total viable cells were used for analysis.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5 Effect of nicotine on the P. gingivalis LPS-induced protein expression of pro-inflammatory mediators in HUVECs. HUVECs were stimulated by P. gingivalis LPS in the presence or absence of nicotine (10 uM-10 mM) for 4, 24, and 72 h. After stimulation, the surface expression levels of ICAM-1 (A), VCAM-1 (B), and E-selectin (C) were measured by flow cytometry, and the quantity of MCP-1 (D) and IL-8 (E) in conditioned media was measured by ELISA. Each value represents mean +-SD of three independent assays. Non-stimulated HUVECs were used as a control. The protein expression levels of pro-inflammatory mediators were not analyzed after stimulation with 10-mM nicotine for 24 and 72 h because the cells were not viable. * - significantly different between groups, p