Clone:
REA472
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Alternative names:
6M21, Ly89, PIRA1, Pir, GP91, LIR-3, LILRB3, PIR-B

Extended validation for PIR-A/B 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 REA472
326414-
6C1++
10-1-PIR-
404127-
REA899-
Cells were incubated with an excess of purified unconjugated PIR-A/B (REA472) 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 PIR-A/B. C57BL6/6 splenocytes were stained with PIR-A/B antibodies and with a suitable counterstaining. As a control, PIR-A/B 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 PIR-A/B. C57BL6/6 splenocytes were stained with PIR-A/B antibodies and with a suitable counterstaining. As a control, PIR-A/B 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 PIR-A/B. C57BL6/6 splenocytes were stained with PIR-A/B antibodies and with a suitable counterstaining. As a control, PIR-A/B 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 PIR-A/B. C57BL6/6 splenocytes were stained with PIR-A/B antibodies and with a suitable counterstaining. As a control, PIR-A/B 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 PIR-A/B (REA472). 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 PIR-A/B (REA472). 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 PIR-A/B (REA472). 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 PIR-A/B Antibody, anti-mouse, REAfinity™

Overview

Clone REA472 recognizes both isoforms of the mouse paired immunoglobulin-like receptor (PIR-A and PIR-B) antigen, a single-pass type I membrane protein which is also known as leukocyte immunoglobulin-like receptor 3 (LIR-3). Cell surface expression of PIR molecules on myeloid and B lineage cells increase with cellular differentiation and activation. PIR molecules are also expressed on dendritic cells, monocytes/macrophages, and mast cells in varying levels, but not on T cells and NK cells. PIR-A/B is a receptor for class I MHC antigens and down-regulates antigen-induced B cell–activation by recruiting phosphatases to its immunoreceptor tyrosine-based inhibitor motifs (ITIM).
Additional information: Clone REA472 displays negligible binding to Fc receptors.

Alternative names

6M21, Ly89, PIRA1, Pir, GP91, LIR-3, LILRB3, PIR-B

Detailed product information

Technical specifications

CloneREA472
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenPIR-A/B
Alternative names of antigen6M21, Ly89, PIRA1, Pir, GP91, LIR-3, LILRB3, PIR-B
Molecular mass of antigen [kDa]83(average of subunits)
Distribution of antigenB cells, dendritic cells, macrophages, mast cells, monocytes
Entrez Gene ID18722, 18733
RRIDAB_2653200, AB_2653201, AB_2653202, AB_2653203, AB_2653204, AB_2653205, AB_2653206, AB_2653199

References for PIR-A/B Antibody, anti-mouse, REAfinity™

Publications

  1. Kubagawa, H. et al. (1999) Biochemical nature and cellular distribution of the paired immunoglobulin-like receptors, PIR-A and PIR-B. J. Exp. Med. 189(2): 309-318
  2. Bléry, M. et al. (1998) The paired Ig-like receptor PIR-B is an inhibitory receptor that recruits the protein-tyrosine phosphatase SHP-1. Proc. Natl. Acad. Sci. U.S.A. 95(5): 2446-2451
  3. Kubagawa, H. et al. (1997) A novel pair of immunoglobulin-like receptors expressed by B cells and myeloid cells. Proc. Natl. Acad. Sci. U.S.A. 94(10): 5261-5266

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