Clone:
REA280
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
SELE, Elam, ESEL, LECAM2, ELAM-1, E-selectin

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

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA280
HAE-1f++
HCD62E++
68-5H11++
P2H3+
Cells were incubated with an excess of purified unconjugated CD62E (REA280) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD62E. Human umbelical vein endithelial cells (Huvec) were stimulated with 25ng/mL TNFα a for 6 hours. Cells were stained with CD62E antibodies and plotted against the side scatter. 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.
View details
Flow cytometric comparison of different clones for CD62E. Human umbelical vein endithelial cells (Huvec) were stimulated with 25ng/mL TNFα a for 6 hours. Cells were stained with CD62E antibodies and plotted against the side scatter. 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.
View details
Flow cytometric comparison of different clones for CD62E. Human umbelical vein endithelial cells (Huvec) were stimulated with 25ng/mL TNFα a for 6 hours. Cells were stained with CD62E antibodies and plotted against the side scatter. 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.
View details
Flow cytometric comparison of different clones for CD62E. Human umbelical vein endithelial cells (Huvec) were stimulated with 25ng/mL TNFα a for 6 hours. Cells were stained with CD62E antibodies and plotted against the side scatter. 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.
View details
Flow cytometric comparison of different clones for CD62E. Human umbelical vein endithelial cells (Huvec) were stimulated with 25ng/mL TNFα a for 6 hours. Cells were stained with CD62E antibodies and plotted against the side scatter. 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. Human umbelical vein endithelial cells (Huvec) were stimulated with 25ng/mL TNFα a for 6 hours. Cells were stained with CD62E antibodies and plotted against the side scatter. 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
View details
Comparison of staining pattern on non-fixed and fixed cells using CD62E (REA280). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD62E (REA280). 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 (REA280). 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-human, REAfinity™

Overview

Clone REA280 recognizes the CD62 antigen-like family member E (CD62E) antigen, a 115 kDa single-pass type I membrane protein which is also known as E-selectin, endothelial-leukocyte adhesion molecule 1 (ELAM-1), or leukocyte-endothelial cell adhesion molecule 2 (LECAM2). CD62E is a cell adhesion molecule highly expressed only on endothelial cells activated by cytokines. It is not stored in the cell and has to be transcribed, translated, and transported to the cell surface. The production of CD62E is stimulated by the expression of CD62P which is stimulated by tumor necrosis factor α, and it can also be stimulated by interleukin-1 and lipopolysaccharide. CD62E recognizes and binds to sialylated carbohydrates present on the surface proteins of certain leukocytes. CD62E ligands are expressed by neutrophils, monocytes, eosinophils, memory-effector T-like lymphocytes, and natural killer cells. Each of these cell types is found in acute and chronic inflammatory sites in association with expression of CD62E, thus implicating CD62E in the recruitment of these cells to such inflammatory sites.
Additional information: Clone REA280 displays negligible binding to Fc receptors.

Alternative names

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

Detailed product information

Technical specifications

CloneREA280
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD62E
Alternative names of antigenSELE, Elam, ESEL, LECAM2, ELAM-1, E-selectin
Molecular mass of antigen [kDa]65
Distribution of antigenmonocytes, NK cells, lymphocytes, eosinophils, neutrophils
Entrez Gene ID6401
RRIDAB_2658845, AB_2658846, AB_2658847, AB_2658848, AB_2658849, AB_2658850, AB_2658851, AB_2658844

References for CD62E Antibody, anti-human, REAfinity™

Publications

  1. Hession, C. et al. (1990) Endothelial leukocyte adhesion molecule 1: direct expression cloning and functional interactions. Proc. Natl. Acad. Sci. U.S.A. 87(5): 1673-1677
  2. Somers, W. S. et al. (2000) Insights into the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin bound to SLe(X) and PSGL-1. Cell 103(3): 467-479
  3. Walzog, B. et al. (2000) Adhesion molecules: the path to a new understanding of acute inflammation. News Physiol. Sci. 15: 107-113

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