Clone:
REA1071
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
5H9, MRP-1, Tetraspanin-29 (Tspan-29), TSPAN-29, p24, MIC3, Tspan29, ORF, GIG2

Extended validation for CD9 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 REA1071
SN4 C3-3A2++
HI9a+
M-L13++
REAL500++
Cells were incubated with an excess of purified unconjugated CD9 (REA1071) 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 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.
View details
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.
View details
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.
View details
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.
View details
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
View details
Comparison of staining pattern on non-fixed and fixed cells using CD9 (REA1071). 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 CD9 (REA1071). 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 (REA1071). 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, REAfinity™

Overview

Clone REA1071 recognizes the human CD9 antigen, a 25 kDa cell surface glycoprotein which is also known as tetraspanin-29 (Tspan-29). CD9 belongs to the tetraspanin family of integral proteins, which are characterized 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 costimulatory signal during T cell activation.
Additional information: Clone REA1071 displays negligible binding to Fc receptors.

Alternative names

5H9, MRP-1, Tetraspanin-29 (Tspan-29), TSPAN-29, p24, MIC3, Tspan29, ORF, GIG2

Detailed product information

Technical specifications

CloneREA1071
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD9
Alternative names of antigen5H9, MRP-1, Tetraspanin-29 (Tspan-29), TSPAN-29, p24, MIC3, Tspan29, ORF, GIG2
Molecular mass of antigen [kDa]25
Distribution of antigenB cells, epithelial cells, fibroblasts, granulocytes, macrophages, platelets, T cells, eosinophils, pre B cells, tumor
Entrez Gene ID928
RRIDAB_2733658, AB_2733844, AB_2733845, AB_2733405, AB_2733406, AB_2733954, AB_2733955, AB_2734094, AB_2734095, AB_2732986, AB_2732987, AB_2733252, AB_2733253, AB_2733145, AB_2733146, AB_2733043, AB_2733044, AB_2733538, AB_2733539, AB_2733657

Resources for CD9 Antibody, anti-human, REAfinity™

Certificates

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References for CD9 Antibody, anti-human, REAfinity™

Publications

  1. 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
  2. Jones, E. L. et al. (2011) Tetraspanins in cellular immunity. Biochem. Soc. T. 39(2): 506-511
  3. 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

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