Clone:
REA802
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
PDCD1, Ly101, PD1, Pdc1

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 clonesOverlap 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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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
View details
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.
View details
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

CloneREA802
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD279 (PD1)
Alternative names of antigenPDCD1, Ly101, PD1, Pdc1
Molecular mass of antigen [kDa]29
Distribution of antigenB cells, monocytes, T cells, thymocytes, spleen
Entrez Gene ID18566
RRIDAB_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

  1. 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
  2. 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
  3. Hirano, F. et al. (2005) Blockade of B7-H1 and PD-1 by monoclonal antibodies potentiates cancer therapeutic immunitiy. Cancer Res. 65: 1089-1096
  4. Tsushima, F. et al. (2007) Interaction between B7-H1 and PD-1 determines initiation and reversal of T cell anergy. Blood 110: 180-185

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