CD146 (LSEC) Antibody, anti-mouse

Name: CD146 (LSEC) Antibody, anti-mouse
Availability: InStock

CD146 (LSEC) Antibody, anti-mouseExtended Validation

Clone:

ME-9F1

Type of antibody:

Primary antibodies

Isotype:

rat IgG2aκ

Applications:

FC, MICS, IF, IHC

Alternative names:

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

Flow cytometry applications forCD146 (LSEC) Antibody, anti-mouse

APC
Biotin
FITC
PE
PerCP-Vio 700
VioBlue

Conjugate:

APC

Recommended dilution:

1:10

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-102-846, 130-102-277

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:10)

Description:

Mouse liver cells were stained with CD146 (LSEC) antibodies. Cells stained with CD146 (LSEC)-Biotin were also stained with Anti-Biotin-APC and analyzed by flow cytometry. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

MICS (MACSima Imaging Cyclic Staining) applications forCD146 (LSEC) Antibody, anti-mouse

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:10

Fixation:

PFA

Tested:

during development

Species tested:

mouse

Products used:

130-102-846, 130-102-277

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:10

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-102-846, 130-102-277

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD146 (LSEC) Antibody, anti-mouse

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 clones	Overlap in epitope recognition with ME-9F1
REA1064	++
Cells were incubated with an excess of purified unconjugated CD146 (LSEC) (ME-9F1) 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.

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.

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

Comparison of staining pattern on non-fixed and fixed cells using CD146 (LSEC) (ME-9F1). 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) (ME-9F1). 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

Overview

The CD146 (LSEC) antibody (clone ME-9F1) binds to the CD146 antigen, which is expressed on mouse endothelial cells, including liver sinusoidal endothelial cells (LSECs), smooth muscle cells, and the basal membrane.

¹

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.

²

Alternative names

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

Detailed product information

Technical specifications

Clone	ME-9F1
Clonality	monoclonal
Isotype	rat IgG2aκ
Isotype control	Isotype Control Antibody, rat IgG2a
Host	rat
Type of antibody	Primary antibodies
Species	mouse
Antigen	CD146 (LSEC)
Alternative names of antigen	MCAM, 1-gicerin, CD149, MUC18, S-endo, s-gicerin, Endo-CAM, Mel-CAM
Molecular mass of antigen [kDa]	69
Distribution of antigen	dendritic cells, endothelial cells, epithelial cells, fibroblasts, T cells, stem cells, mesenchymal stem cells, ES and iPS cells, bone marrow, smooth muscle
Entrez Gene ID	84004
RRID	AB_2661096, AB_2661097, AB_2661098, AB_2661099, AB_2661100, AB_2661101, AB_2661102, AB_2661103, AB_2661104, AB_2661105, AB_2661106, AB_2661095

Resources for CD146 (LSEC) Antibody, anti-mouse

Certificates

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References for CD146 (LSEC) Antibody, anti-mouse

Publications

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
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

Applications

Extended validation