Clone:
REA990
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
ICOSL, B7-H2, B7RP-1, B7h, ICOSLG, LICOS, Ly115l, GL50

Extended validation for CD275 (B7-H2) 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 REA990
HK5.3++
Cells were incubated with an excess of purified unconjugated CD275 (B7-H2) (REA990) 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 CD275 (B7-H2). Mix of unseparated C57BL/6 Mouse Splenocytes and isolated B-cells stored overnight were stained with CD275 (B7-H2) antibodies and with a suitable counterstaining. As a control, CD275 (B7-H2) 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 CD275 (B7-H2). Mix of unseparated C57BL/6 Mouse Splenocytes and isolated B-cells stored overnight were stained with CD275 (B7-H2) antibodies and with a suitable counterstaining. As a control, CD275 (B7-H2) 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 CD275 (B7-H2). Mix of unseparated C57BL/6 Mouse Splenocytes and isolated B-cells stored overnight were stained with CD275 (B7-H2) antibodies and with a suitable counterstaining. As a control, CD275 (B7-H2) 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 CD275 (B7-H2). Mix of unseparated C57BL/6 Mouse Splenocytes and isolated B-cells stored overnight were stained with CD275 (B7-H2) antibodies and with a suitable counterstaining. As a control, CD275 (B7-H2) 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 CD275 (B7-H2). Mix of unseparated C57BL/6 Mouse Splenocytes and isolated B-cells stored overnight were stained with CD275 (B7-H2) antibodies and with a suitable counterstaining. As a control, CD275 (B7-H2) 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 CD275 (B7-H2). Mix of unseparated C57BL/6 Mouse Splenocytes and isolated B-cells stored overnight were stained with CD275 (B7-H2) antibodies and with a suitable counterstaining. As a control, CD275 (B7-H2) 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 CD275 (B7-H2) (REA990). 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 CD275 (B7-H2) (REA990). 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 CD275 (B7-H2) (REA990). 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 CD275 (B7-H2) Antibody, anti-mouse, REAfinity™

Overview

Clone REA990 recognises the mouse CD275 antigen, which is also known as B7-H2 or ICOSL. CD275 is a type I membrane protein and belongs to the Ig superfamily, B7 family of co-stimulatory molecules. Expression of CD275 is found on antigen presenting cells such as B cells, dendritic cells, and monocytes. CD275 co-stimulates T cells upon binding to inducible T cell co-stimulator (ICOS), which is expressed on activated T cells. In mouse, CD275 binds only ICOS, whereas in humans it can interact with other T cell co-stimulatory molecules, CD28 and CTLA-4. To appropriately regulate the activation of immune response, the ICOS-CD275 interaction can also lead to downregulation of CD275 expression which allows prevention of hyperreaction of T cells.
Additional information: Clone REA990 displays negligible binding to Fc receptors.

Alternative names

ICOSL, B7-H2, B7RP-1, B7h, ICOSLG, LICOS, Ly115l, GL50

Detailed product information

Technical specifications

CloneREA990
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD275 (B7-H2)
Alternative names of antigenICOSL, B7-H2, B7RP-1, B7h, ICOSLG, LICOS, Ly115l, GL50
Molecular mass of antigen [kDa]31
Distribution of antigendendritic cells, macrophages, monocytes, B cells, antigen-presenting cells
Entrez Gene ID50723
RRIDAB_2727547, AB_2727507, AB_2727548, AB_2727551, AB_2727509, AB_2727549, AB_2727550, AB_2727508, AB_2727546, AB_2727506

Resources for CD275 (B7-H2) Antibody, anti-mouse, REAfinity™

Certificates

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to search for Certificates of Analysis (CoA) by lot number.

References for CD275 (B7-H2) Antibody, anti-mouse, REAfinity™

Publications

  1. Watanabe, M. et al. (2008) Down-regulation of ICOS ligand by interaction with ICOS functions as a regulatory mechanism for immune responses. J. Immunol. 180: 5222-5234
  2. Yao, S. et al. (2011) B7-H2 is a costimulatory ligand for CD28 in human. Immunity 34(5): 729-740

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