CD279 (PD1) Antibody, anti-mouse, REAfinity™

Name: CD279 (PD1) Antibody, anti-mouse, REAfinity™
Availability: InStock

CD279 (PD1) Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA802

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

PDCD1, Ly101, PD1, Pdc1

Flow cytometry applications forCD279 (PD1) Antibody, anti-mouse, REAfinity™

APC
Biotin
FITC
PE
PerCP-Vio 700

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:

130-111-954, 130-111-801

Protocols:

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

Description:

Mouse thymocytes from 10 week old BALB/c mice were stained with CD279 (PD1) antibodies or with the corresponding REA Control antibodies as well as with CD8a and CD4 antibodies. CD8a^– and CD8a^dim cells were pre-gated. Cells were analyzed by flow cytometry using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forCD279 (PD1) Antibody, anti-mouse, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-111-954, 130-111-801

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD279 (PD1) 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 REA802
29F.1A12	+
RMP1-14	-
RMP1-30	++
J43	++
766104	-
HA2-7B1	++
REAL269	++
Cells were incubated with an excess of purified unconjugated CD279 (PD1) (REA802) 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 forCD279 (PD1). Balb/c thymocytes were stained with CD279 (PD1) antibodies and with a suitable counterstaining. As a control, CD279 (PD1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cells were pregated on CD8a negative cells. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones forCD279 (PD1). Balb/c thymocytes were stained with CD279 (PD1) antibodies and with a suitable counterstaining. As a control, CD279 (PD1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cells were pregated on CD8a negative cells. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones forCD279 (PD1). Balb/c thymocytes were stained with CD279 (PD1) antibodies and with a suitable counterstaining. As a control, CD279 (PD1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cells were pregated on CD8a negative cells. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones forCD279 (PD1). Balb/c thymocytes were stained with CD279 (PD1) antibodies and with a suitable counterstaining. As a control, CD279 (PD1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cells were pregated on CD8a negative cells. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones forCD279 (PD1). Balb/c thymocytes were stained with CD279 (PD1) antibodies and with a suitable counterstaining. As a control, CD279 (PD1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cells were pregated on CD8a negative cells. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones forCD279 (PD1). Balb/c thymocytes were stained with CD279 (PD1) antibodies and with a suitable counterstaining. As a control, CD279 (PD1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cells were pregated on CD8a negative cells. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones forCD279 (PD1). Balb/c thymocytes were stained with CD279 (PD1) antibodies and with a suitable counterstaining. As a control, CD279 (PD1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cells were pregated on CD8a negative cells. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones forCD279 (PD1). Balb/c thymocytes were stained with CD279 (PD1) antibodies and with a suitable counterstaining. As a control, CD279 (PD1) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cells were pregated on CD8a negative cells. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. 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 CD279 (PD1) (REA802). 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 CD279 (PD1) (REA802). 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 CD279 (PD1) Antibody, anti-mouse, REAfinity™

Overview

Clone REA802 recognizes the mouse CD279 antigen, also known as PD1. CD279 is a member of the Ig superfamily expressed mainly on activated T and B lymphocytes. Expression is induced on activated myeloid cells as well. CD279 is thought to be involved in lymphocyte clonal selection and plays a key role in peripheral tolerance and autoimmune disease in mice. The ligands of CD279, PDL1 (B7-H1) and PDL2 (B7-DC), belong to the B7 immunoglobulin superfamily.

Additional information: Clone REA802 displays negligible binding to Fc receptors.

Alternative names

PDCD1, Ly101, PD1, Pdc1

Detailed product information

Technical specifications

Clone	REA802
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	CD279 (PD1)
Alternative names of antigen	PDCD1, Ly101, PD1, Pdc1
Molecular mass of antigen [kDa]	29
Distribution of antigen	B cells, monocytes, T cells, thymocytes, spleen
Entrez Gene ID	18566
RRID	AB_2656932, AB_2656933, AB_2656934, AB_2656935, AB_2656936, AB_2656937, AB_2656938, AB_2656939, AB_2656940, AB_2656931

Resources for CD279 (PD1) Antibody, anti-mouse, REAfinity™

Certificates

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References for CD279 (PD1) Antibody, anti-mouse, REAfinity™

Publications

Iwai, Y. et al. (2002) Involvement of PD-L1 on tumor cells in the escape from host immune system and tumor immunotherapy by PD-L1 blockade. Proc. Natl. Acad. Sci. U.S.A. 99: 12293-12297
Salama, A. D. et al. (2003) Critical role of the programmed death-1 (PD-1) pathway in regulation of experimental autoimmune encephalomyelitis. J. Exp. Med. 198: 71-78
Hirano, F. et al. (2005) Blockade of B7-H1 and PD-1 by monoclonal antibodies potentiates cancer therapeutic immunitiy. Cancer Res. 65: 1089-1096
Tsushima, F. et al. (2007) Interaction between B7-H1 and PD-1 determines initiation and reversal of T cell anergy. Blood 110: 180-185

Applications

Extended validation