Clone:
REA1064
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
MCAM, 1-gicerin, CD149, MUC18, S-endo, s-gicerin, Endo-CAM, Mel-CAM

Extended validation for CD146 (LSEC) Antibody, anti-mouse, 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 REA1064
ME-9F1++
Cells were incubated with an excess of purified unconjugated CD146 (LSEC) (REA1064) 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 CD146 (LSEC). Splenocytes from CD57BL/6 mice were stained with CD146 (LSEC) antibodies and with a suitable counterstaining. As a control, CD146 (LSEC) 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 CD146 (LSEC). Splenocytes from CD57BL/6 mice were stained with CD146 (LSEC) antibodies and with a suitable counterstaining. As a control, CD146 (LSEC) 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 CD146 (LSEC). Splenocytes from CD57BL/6 mice were stained with CD146 (LSEC) antibodies and with a suitable counterstaining. As a control, CD146 (LSEC) 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 CD146 (LSEC). Splenocytes from CD57BL/6 mice were stained with CD146 (LSEC) antibodies and with a suitable counterstaining. As a control, CD146 (LSEC) 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 CD146 (LSEC). Splenocytes from CD57BL/6 mice were stained with CD146 (LSEC) antibodies and with a suitable counterstaining. As a control, CD146 (LSEC) 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 CD146 (LSEC) (REA1064). 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 CD146 (LSEC) (REA1064). 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 CD146 (LSEC) (REA1064). 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 CD146 (LSEC) Antibody, anti-mouse, REAfinity™

Overview

Clone REA1064 recognizes the mouse CD146 antigen, which is expressed on mouse endothelial cells, including liver sinusoidal endothelial cells (LSECs), smooth muscle cells, and the basal membrane. LSECs are microvascular endothelial cells lining the hepatic sinusoidal wall. Their strategic positioning favors a tight interaction with lymphocytes migrating through the liver. They possess a high capacity for antigen uptake and processing. However, in contrast to professional antigen-presenting cells, e.g. dendritic cells, they express only low levels of costimulatory molecules.
Additional information: Clone REA1064 displays negligible binding to Fc receptors.

Alternative names

MCAM, 1-gicerin, CD149, MUC18, S-endo, s-gicerin, Endo-CAM, Mel-CAM

Detailed product information

Technical specifications

CloneREA1064
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD146 (LSEC)
Alternative names of antigenMCAM, 1-gicerin, CD149, MUC18, S-endo, s-gicerin, Endo-CAM, Mel-CAM
Molecular mass of antigen [kDa]69
Distribution of antigendendritic cells, endothelial cells, epithelial cells, fibroblasts, stem cells, T cells, ES and iPS cells, mesenchymal stem cells
Entrez Gene ID84004
RRIDAB_2751472, AB_2751507, AB_2751473, AB_2751508, AB_2751474, AB_2751509, AB_2751475, AB_2751510, AB_2751511, AB_2751506

References for CD146 (LSEC) Antibody, anti-mouse, REAfinity™

Publications

  1. Harder, R. et al. (1991) Dissection of murine lymphocyte-endothelial cell interaction mechanisms by SV-40-transformed mouse endothelial cell lines: novel mechanisms mediating basal binding, and alpha 4-integrin-dependent cytokine-induced adhesion. Exp. Cell Res. 197: 259-267
  2. Diehl, L. et al. (2008)
    Tolerogenic maturation of liver sinusoidal endothelial cells promotes B7-homolog 1-dependent CD8
    +
    T cell tolerance.
    Hepatology 47: 296-305

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