Clone:
REA615
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
L-selectin, LAM-1, LECAM-1, Leu-8, TQ1, gp90-MEL

Extended validation for CD62L 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 REA615
DREG-56-
SK11++
LT-TD180-
1H3++
145/15++
REAL163++
Cells were incubated with an excess of purified unconjugated CD62L (REA615) 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 CD62L. Human peripheral blood mononuclear cells (PBMCs) were stained with CD62L antibodies and with a suitable counterstaining. As a control, CD62L 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 CD62L. Human peripheral blood mononuclear cells (PBMCs) were stained with CD62L antibodies and with a suitable counterstaining. As a control, CD62L 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 CD62L. Human peripheral blood mononuclear cells (PBMCs) were stained with CD62L antibodies and with a suitable counterstaining. As a control, CD62L 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 CD62L. Human peripheral blood mononuclear cells (PBMCs) were stained with CD62L antibodies and with a suitable counterstaining. As a control, CD62L 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 CD62L. Human peripheral blood mononuclear cells (PBMCs) were stained with CD62L antibodies and with a suitable counterstaining. As a control, CD62L 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 CD62L. Human peripheral blood mononuclear cells (PBMCs) were stained with CD62L antibodies and with a suitable counterstaining. As a control, CD62L 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 CD62L. Human peripheral blood mononuclear cells (PBMCs) were stained with CD62L antibodies and with a suitable counterstaining. As a control, CD62L 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 CD62L. Human peripheral blood mononuclear cells (PBMCs) were stained with CD62L antibodies and with a suitable counterstaining. As a control, CD62L 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 CD62L. Human peripheral blood mononuclear cells (PBMCs) were stained with CD62L antibodies and with a suitable counterstaining. As a control, CD62L 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 CD62L (REA615). 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 CD62L (REA615). 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 CD62L (REA615). 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 CD62L Antibody, anti-human, REAfinity™

Overview

Clone REA615 recognizes the human CD62L antigen, a 74 kDa single-pass type I membrane protein, which is also known as L-selectin, LECAM-1, or LAM-1. CD62L is a member of the selectin family of cell surface molecules and binds a series of glycoproteins including CD34, GlyCAM-1, and MAdCAM-1. Most hematopoietic cells express CD62L, including peripheral blood B cells, T cells, monocytes, granulocytes, thymocytes, and some myeloid cells from bone marrow. Expression on resting naive, central memory, and some effector memory T cells regulates their migration via endothelial venules to peripheral lymph nodes and Peyer’s patches. The CD62L antigen also contributes to the recruitment of leukocytes from the blood to areas of inflammation. Disruption of CD62L expression has detrimental effects on T cell migration and immune responses. Antigenic activation in the lymph node leads first to rapid CD62L shedding by protease cleavage and then to transcriptional suppression.
Always use fresh material for immunofluorescent staining of CD62L
+
cells. For optimal results, the cells should not be older than 8–12 hours. Keep cells continuously cold. CD62L-expression may be rapidly lost due to shedding.
Additional information: Clone REA615 displays negligible binding to Fc receptors.

Alternative names

L-selectin, LAM-1, LECAM-1, Leu-8, TQ1, gp90-MEL

Detailed product information

Technical specifications

CloneREA615
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, non-human primate
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
,
cynomolgus monkey (
Macaca fascicularis
)
, baboon
AntigenCD62L
Alternative names of antigenL-selectin, LAM-1, LECAM-1, Leu-8, TQ1, gp90-MEL
Molecular mass of antigen [kDa]38
Distribution of antigenB cells, granulocytes, Langerhans cells, lymphocytes, monocytes, NK cells, red blood cells, T cells, eosinophils, neutrophils, thymocytes, bone marrow, spleen
Entrez Gene ID6402
RRIDAB_2733829, AB_2751219, AB_2751153, AB_2819417, AB_2733828

References for CD62L Antibody, anti-human, REAfinity™

Publications

  1. Ivetic, A. (2013) Signals regulating L-selectin-dependent leucocyte adhesion and transmigration. Int. J. Biochem. Cell Biol. 45(3): 550-555
  2. McEver, R. P. (2015) Selectins: initiators of leucocyte adhesion and signalling at the vascular wall. Cardiovasc. Res. 107(3): 331-339
  3. Telen, M. J. (2014) Cellular adhesion and the endothelium: E-selectin, L-selectin, and pan-selectin inhibitors. Hematol. Oncol. Clin. North Am. 28(2): 341-354

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