CD101 Antibody, anti-human

Name: CD101 Antibody, anti-human
Availability: InStock

CD101 Antibody, anti-humanExtended Validation

Clone:

BB27

Type of antibody:

Primary antibodies

Isotype:

mouse IgG1κ

Applications:

FC, MICS, IF, IHC, ICC

Alternative names:

EWI-101, IGSF2, V7, p126

Flow cytometry applications forCD101 Antibody, anti-human

APC
Biotin
PE
PE-Vio 770
Vio Bright FITC

Conjugate:

APC

Recommended dilution:

1:11

Remark:

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

Tested:

QC tested

Products used:

130-095-132

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:11)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD101 antibodies as well as with CD4 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forCD101 Antibody, anti-human

APC
PE

Conjugate:

APC

Recommended dilution:

1:11

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-095-132

Compatible applications:

IF, IHC, ICC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:11

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-095-132

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD101 Antibody, anti-human

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 BB27
REA954	++
V7.1	-
Cells were incubated with an excess of purified unconjugated CD101 (BB27) 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 CD101. Human peripheral blood mononuclear cells (PBMCs) were stained with CD101 antibodies and with a suitable counterstaining. As a control, CD101 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 CD101. Human peripheral blood mononuclear cells (PBMCs) were stained with CD101 antibodies and with a suitable counterstaining. As a control, CD101 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 CD101. Human peripheral blood mononuclear cells (PBMCs) were stained with CD101 antibodies and with a suitable counterstaining. As a control, CD101 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 CD101. Human peripheral blood mononuclear cells (PBMCs) were stained with CD101 antibodies and with a suitable counterstaining. As a control, CD101 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 CD101. Human peripheral blood mononuclear cells (PBMCs) were stained with CD101 antibodies and with a suitable counterstaining. As a control, CD101 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 CD101 (BB27). 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 CD101 (BB27). 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 CD101 Antibody, anti-human

Overview

Clone BB27 recognizes the CD101 antigen, also known as V7 or IGSF2, which is a 140 kDa type I transmembrane glycoprotein of the Ig gene superfamily. CD101 is expressed on monocytes, granulocytes, and subsets of dendritic cells (DCs). In addition, CD101 is expressed on certain subpopulations of T cells, in particular about 5–30% of FoxP3

⁺

Treg cells in peripheral blood.

The interaction of CD101 with the antibody BB27 has been shown to inhibit T cell activation by allogeneic DCs or CD3 antibodies.

Alternative names

EWI-101, IGSF2, V7, p126

Detailed product information

Technical specifications

Clone	BB27
Clonality	monoclonal
Isotype	mouse IgG1κ
Isotype control	Isotype Control Antibody, mouse IgG1
Host	mouse
Type of antibody	Primary antibodies
Species	human
Antigen	CD101
Alternative names of antigen	EWI-101, IGSF2, V7, p126
Molecular mass of antigen [kDa]	113
Distribution of antigen	dendritic cells, granulocytes, Langerhans cells, monocytes, red blood cells, T cells, lymphocytes, leukemia cells, thymocytes, lung, small intestine, spleen
Entrez Gene ID	9398
RRID	AB_2661056, AB_10827694, AB_10829614, AB_2661057, AB_2661058, AB_10828923, AB_2661055

Resources for CD101 Antibody, anti-human

Certificates

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References for CD101 Antibody, anti-human

Publications

Rivas, A. et al. (1995) V7, a novel leukocyte surface protein that participates in T cell activation. J Immunol 154: 4423-4433
Soares, L. R. et al. (1998)
V7 (CD101) ligation inhibits TCR/CD3-induced IL-2 production by blocking Ca
²⁺
flux and nuclear factor of activated T cell nucleartranslocation.
J Immunol 161: 209-217
Bouloc, A. et al. (2000) Triggering CD101 molecule on human cutaneous dendritic cells inhibits T cell proliferation via IL-10 production. Eur. J. Immunol. 30: 3132-3139
Bagot, M. et al. (1997) CD101 is expressed by skin dendritic cells: role in T-lymphocyte activation. Tissue Antigens 50: 439-448

Applications

Extended validation