CD62E Antibody, anti-mouse, REAfinity™

Name: CD62E Antibody, anti-mouse, REAfinity™
Availability: InStock

CD62E Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA369

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

SELE, E-selectin, ELAM-1, Elam, LECAM2

Flow cytometry applications forCD62E Antibody, anti-mouse, REAfinity™

APC
Biotin
PE

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-125-992

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Mouse bEnd.3 cells were either left unstimulated (middle peak) or stimulated with 20 ng/mL TNF-α for 5 hours. Cells were then stained with CD62E antibodies (right peak) or with the corresponding REA Control (S) antibodies (left peak) and analyzed by flow cytometry using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Extended validation for CD62E Antibody, anti-mouse, REAfinity™

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

Flow cytometric comparison of different clones for CD62E. C57BL/6 Mouse splenocytes were stained with CD62E antibodies and with a suitable counterstaining. As a control, CD62E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD62E. C57BL/6 Mouse splenocytes were stained with CD62E antibodies and with a suitable counterstaining. As a control, CD62E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD62E. C57BL/6 Mouse splenocytes were stained with CD62E antibodies and with a suitable counterstaining. As a control, CD62E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD62E. C57BL/6 Mouse splenocytes were stained with CD62E antibodies and with a suitable counterstaining. As a control, CD62E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD62E. C57BL/6 Mouse splenocytes were stained with CD62E antibodies and with a suitable counterstaining. As a control, CD62E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD62E. C57BL/6 Mouse splenocytes were stained with CD62E antibodies and with a suitable counterstaining. As a control, CD62E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD62E. C57BL/6 Mouse splenocytes were stained with CD62E antibodies and with a suitable counterstaining. As a control, CD62E antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Fixation data

To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.

No fixation

Post fixation

Comparison of staining pattern on non-fixed and fixed cells using CD62E (REA369). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using CD62E (REA369). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using CD62E (REA369). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using CD62E (REA369). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD62E Antibody, anti-mouse, REAfinity™

Overview

Clone REA369 recognizes the mouse CD62E antigen, a single-pass type I membrane glycoprotein also known as E-selectin. CD62E is expressed on endothelial cells and plays a critical role in the recruitment of the appropriate blood cell into specific local sites. It mediates the adhesion of blood neutrophils in cytokine-activated endothelium through interaction with P-selectin glycoprotein ligand 1, which promotes slow rolling and adhesion. CD62E is transcriptionally induced in response to inflammatory cytokines, such as IL-1β and TNFα. It has an important role during hematopoietic stem/progenitor cell trafficking to the bone marrow, and for the maintenance of their quiescent phenotype

in vivo

. Because selectins are also constitutively expressed in the endothelium of certain tissues such as bone marrow even in the absence of inflammatory stimulation, there has been a growing appreciation that these receptors may be important for tissue homeostasis. Elevated cellular levels of CD62E have been documented in various diseases associated with a pro-inflammatory condition, including diabetes, atherosclerosis, rheumatoid arthritis, and cancer. Conversely, therapeutic strategies, which reduce vascular injury, result in significant down-regulation of CD62E gene expression.

Additional information: Clone REA369 displays negligible binding to Fc receptors.

Alternative names

SELE, E-selectin, ELAM-1, Elam, LECAM2

Detailed product information

Technical specifications

Clone	REA369
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody, human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	mouse
Antigen	CD62E
Alternative names of antigen	SELE, E-selectin, ELAM-1, Elam, LECAM2
Molecular mass of antigen [kDa]	65
Distribution of antigen	endothelial cells, bone marrow
Entrez Gene ID	20339
RRID	AB_2876943, AB_2658842, AB_2658843, AB_2751781

Resources for CD62E Antibody, anti-mouse, REAfinity™

Certificates

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References for CD62E Antibody, anti-mouse, REAfinity™

Publications

Katayama, Y. et al. (2013) Selective E-selectin ligands. Blood 122(24): 3858-3860
Weller, A. et al. (1992) Cloning of the mouse endothelial selectins. Expression of both E- and P-selectin is inducible by tumor necrosis factor alpha. J. Biol. Chem. 267(21): 15176-15183
Laviola, L. et al. (2013) TNFα signals via p66(Shc) to induce E-selectin, promote leukocyte transmigration and enhance permeability in human endothelial cells. PLoS One 8(12): e81930

Applications

Extended validation