CD273 (PD-L2) Antibody, anti-mouse, REAfinity™

Name: CD273 (PD-L2) Antibody, anti-mouse, REAfinity™
Availability: InStock

CD273 (PD-L2) Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA986

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

PDCD1LG2, B7-DC, Btdc, PD-L2

Flow cytometry applications forCD273 (PD-L2) Antibody, anti-mouse, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright B515
VioBlue

Conjugate:

APC

Recommended dilution:

1:50

Remark:

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

Tested:

QC tested

Products used:

130-116-767

Protocols:

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

Description:

Isolated CD11c⁺ cells of splenocytes from C57BL/6 mice were stimulated with 20 ng/mL mouse IL-4 for 3 days. Cells were then stained with CD273 (PD-L2) antibodies or with the corresponding REA Control antibodies (left peak), as well as with CD11c antibodies. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

Extended validation for CD273 (PD-L2) 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 clones	Overlap in epitope recognition with REA986
TY25	++
MIH37	++
122	+
MIH37.2.5	++
Cells were incubated with an excess of purified unconjugated CD273 (PD-L2) (REA986) 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 CD273 (PD-L2). PD-L2 transfected cells mixed with mice splenocytes were stained with CD273 (PD-L2) antibodies and with a suitable counterstaining. As a control, CD273 (PD-L2) 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 CD273 (PD-L2). PD-L2 transfected cells mixed with mice splenocytes were stained with CD273 (PD-L2) antibodies and with a suitable counterstaining. As a control, CD273 (PD-L2) 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 CD273 (PD-L2). PD-L2 transfected cells mixed with mice splenocytes were stained with CD273 (PD-L2) antibodies and with a suitable counterstaining. As a control, CD273 (PD-L2) 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 CD273 (PD-L2). PD-L2 transfected cells mixed with mice splenocytes were stained with CD273 (PD-L2) antibodies and with a suitable counterstaining. As a control, CD273 (PD-L2) 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 CD273 (PD-L2). PD-L2 transfected cells mixed with mice splenocytes were stained with CD273 (PD-L2) antibodies and with a suitable counterstaining. As a control, CD273 (PD-L2) 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 CD273 (PD-L2). PD-L2 transfected cells mixed with mice splenocytes were stained with CD273 (PD-L2) antibodies and with a suitable counterstaining. As a control, CD273 (PD-L2) 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 CD273 (PD-L2) (REA986). 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 CD273 (PD-L2) (REA986). 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 CD273 (PD-L2) Antibody, anti-mouse, REAfinity™

Overview

Clone REA986 recognizes CD273, a member of the B7 family of protein ligands. CD273, along with another member of the B7 family (B7-H1) are ligands of T-cell receptor known as programmed death-1 (PD-1). Surface expression of B7-DC is restricted to dendritic cells (DCs) and activated or IFNγ-stimulated DCs and macrophages. CD273/PD-1 interaction is involved in inhibition of T-cell activation, however a co-stimulatory role of CD273, via interaction with receptor other then PD-1 is also reported.

Additional Information: Clone REA986 displays negligible binding to Fc receptors.

Alternative names

PDCD1LG2, B7-DC, Btdc, PD-L2

Detailed product information

Technical specifications

Clone	REA986
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody, human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	mouse
Antigen	CD273 (PD-L2)
Alternative names of antigen	PDCD1LG2, B7-DC, Btdc, PD-L2
Molecular mass of antigen [kDa]	26
Distribution of antigen	dendritic cells, macrophages
Entrez Gene ID	58205
RRID	AB_2904895, AB_2904891, AB_2904899, AB_2904897, AB_2904898, AB_2904892, AB_2904896, AB_2904900, AB_2904893, AB_2904894

Resources for CD273 (PD-L2) Antibody, anti-mouse, REAfinity™

Certificates

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

References for CD273 (PD-L2) Antibody, anti-mouse, REAfinity™

Publications

Tseng, S. Y. et al. (2001)
B7-DC, a new dendritic cell molecule with potent costimulatory properties for T cells.
J. Exp. Med. 193: 839-846
Wang, S. et al. (2003) Molecular modeling and functional mapping of B7-H1 and B7-DC uncouple costimulatory function from PD-1 interaction. J. Exp. Med. 197: 1083
Greenwald, R. J. et al. (2005) The B7 family revisited. Annu. Rev. Immunol. 23: 515-548

Applications

Extended validation