Clone:
REA991
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC
Alternative names:
ICOSLG, B7-H2, B7H2, B7RP-1, GL50, ICOSL, LICOS

Extended validation for CD275 (B7-H2) 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 REA991
MIH12++
2D3-
9F.8A4+
136726++
Cells were incubated with an excess of purified unconjugated CD275 (B7-H2) (REA991) 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). Monocyte derived dendritic cells (MODCs) 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). Monocyte derived dendritic cells (MODCs) 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). Monocyte derived dendritic cells (MODCs) 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). Monocyte derived dendritic cells (MODCs) 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). Monocyte derived dendritic cells (MODCs) 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). Monocyte derived dendritic cells (MODCs) 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). Monocyte derived dendritic cells (MODCs) 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) (REA991). 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) (REA991). 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) (REA991). 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) (REA991). 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) (REA991). 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-human, REAfinity™

Overview

Clone REA991 recognizes the human CD275 antigen, also known as B7-H2, ICOSL, or GL50, a member of the B7 family of co-stimulatory molecules involved in regulating T cell activation and effector functions. It belongs to the Ig superfamily and has a predicted molecular weight of 40 kDa. CD275 is a ligand of inducible co-stimulator (ICOS) expressed on activated T cells. ICOS mediated co-stimulation promotes T helper (Tʜ) cell differentiation via Tʜ2 cytokine production such as interleukin 10 (IL-10) and IL-4. Human CD275 is expressed on B cells, dendritic cells, endothelial cells, renal tubular epithelial cells, activated monocytes, and human airway epithelial cells.
Additional information: Clone REA991 displays negligible binding to Fc receptors.

Alternative names

ICOSLG, B7-H2, B7H2, B7RP-1, GL50, ICOSL, LICOS

Detailed product information

Technical specifications

CloneREA991
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD275 (B7-H2)
Alternative names of antigenICOSLG, B7-H2, B7H2, B7RP-1, GL50, ICOSL, LICOS
Molecular mass of antigen [kDa]31
Distribution of antigenB cells, dendritic cells, endothelial cells, epithelial cells, monocytes, other
Entrez Gene ID23308
RRIDAB_2727687, AB_2727761, AB_2727688, AB_2727762, AB_2727689, AB_2727763, AB_2727690, AB_2727764, AB_2727691, AB_2727765, AB_2727692, AB_2727766, AB_2727693, AB_2727760

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

Certificates

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

Publications

  1. Wang, S. et al. (2000) Costimulation of T cells by B7-H2, a B7-like molecule that binds ICOS. Blood 96(8): 2808-2813
  2. Qian, X. et al. (2006) The ICOS-ligand B7-H2, expessed on human type II alveolar epithelial cells, plays a role in the pulmonary host defense system. Eur. J. Immunol. 36(4): 906-918
  3. Kurosawa, S. et al. (2003) Expression of the costimulatory molecule B7-H2 (inducible costimulator ligand) by human airway epithelial cells. Am. J. Respir. Cell Mol. Biol. 28: 563-573

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