CD9 Antibody, anti-human

Name: CD9 Antibody, anti-human
Availability: InStock

CD9 Antibody, anti-humanExtended Validation

Clone:

SN4 C3-3A2

Type of antibody:

Primary antibodies

Isotype:

mouse IgG1κ

Applications:

FC, MICS, IF, IHC

Alternative names:

BTCC-1, DRAP-27, MIC3, MRP-1, TSPAN-29, Tspan29

Flow cytometry applications forCD9 Antibody, anti-human

APC
APC-Vio 770
Biotin
PE
PE-Vio 770
PerCP-Vio 700

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Reported:

27507971,

Products used:

130-128-101, 130-128-037

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD9 antibodies or with the corresponding isotype control antibodies (left images) and analyzed by flow cytometry using the MACSQuant^® Analyzer. 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 forCD9 Antibody, anti-human

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-128-101, 130-128-037

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-128-101, 130-128-037

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD9 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 SN4 C3-3A2
REA1071	++
HI9a	+
M-L13	++
REAL500	++
Cells were incubated with an excess of purified unconjugated CD9 (SN4 C3-3A2) 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 CD9. Human peripheral blood mononuclear cells (PBMCs) were stained with CD9 antibodies and with a suitable counterstaining. As a control, CD9 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 CD9. Human peripheral blood mononuclear cells (PBMCs) were stained with CD9 antibodies and with a suitable counterstaining. As a control, CD9 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 CD9. Human peripheral blood mononuclear cells (PBMCs) were stained with CD9 antibodies and with a suitable counterstaining. As a control, CD9 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 CD9. Human peripheral blood mononuclear cells (PBMCs) were stained with CD9 antibodies and with a suitable counterstaining. As a control, CD9 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 CD9. Human peripheral blood mononuclear cells (PBMCs) were stained with CD9 antibodies and with a suitable counterstaining. As a control, CD9 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 CD9 (SN4 C3-3A2). 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 CD9 (SN4 C3-3A2). 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 CD9 Antibody, anti-human

Overview

Clone SN4 C3-3A2 recognizes the human CD9 antigen, a 24 kDa cell surface glycoprotein which is also known as tetraspanin-29 (Tspan-29). CD9 belongs to the tetraspanin family of integral proteins, which are characterised by the presence of four conserved transmembrane domains, a small and two large extracellular loops. Expression of CD9 is found on platelets, pre–B cells, fibroblasts, eosinophils, macrophages, basophils, epithelial cells, oocytes, and activated T cells. Further expression is reported on malignant cells and tumor cell lines where CD9 is involved in supressor functions, inhibition of cell invasion, and metastasis. CD9 associates with other tetraspanin members, cell adhesion molecules such as EpCAM, integrin subsets, transmembrane proteins (EWI-2 and EWI-F), choline receptors, and G-proteins. Through diverse interactions or via direct stimulation, CD9 influences various cellular functions including cell adhesion, aggregation, migration, clustering of MHC molecules in dendritic cells (DCs), and provides co-stimulatory signal during T cell activation.

Alternative names

BTCC-1, DRAP-27, MIC3, MRP-1, TSPAN-29, Tspan29

Detailed product information

Technical specifications

Clone	SN4 C3-3A2
Clonality	monoclonal
Isotype	mouse IgG1κ
Isotype control	Isotype Control Antibody, mouse IgG1
Host	mouse
Type of antibody	Primary antibodies
Species	human
Antigen	CD9
Alternative names of antigen	BTCC-1, DRAP-27, MIC3, MRP-1, TSPAN-29, Tspan29
Molecular mass of antigen [kDa]	25
Distribution of antigen	fibroblasts, macrophages, platelets, T cells, eosinophils, basophils
Entrez Gene ID	928
RRID	AB_2857632, AB_2659574, AB_2659575, AB_2659576, AB_2659577, AB_2659578, AB_2659579, AB_2659580, AB_2659581, AB_2905162, AB_2905161, AB_2857657

Resources for CD9 Antibody, anti-human

Certificates

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References for CD9 Antibody, anti-human

Publications

Jones, E. L. et al. (2011) Tetraspanins in cellular immunity. Biochem. Soc. T. 39(2): 506-511
Hemler, M. E. (2003) Tetraspanin proteins mediate cellular penetration, invasion, and fusion events and define a novel type of membrane microdomain. Annu. Rev. Cell Dev. Biol. 19: 397-342
Wang, H. X. et al. (2011) The C-terminal tail of tetraspanin protein CD9 contributes to its function and molecular organization. Cell. Sci. 124: 2702-2710

Applications

Extended validation