Clone:
REAL269
Type of antibody:
Releasable antibodies, Primary antibodies, Recombinant antibodies
Applications:
FC

Extended validation for CD279 (PD1) Antibody, anti-mouse,
REAlease
®

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 REAL269
29F.1A12+
RMP1-14-
RMP1-30++
J43++
766104-
HA2-7B1++
REA802++
Cells were incubated with an excess of purified unconjugated CD279 (PD1) (REAL269) 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) (REAL269). 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) (REAL269). 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) (REAL269). 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,
REAlease
®

Overview

Clone REAL269 is an antibody fragment derived from the full CD279 (PD1) antibody molecule. It displays no binding to Fc receptors. The recombinantly engineered antibody fragments are multimerized to form the REAlease Complex to bind markers with high avidity.
Clone REAL269 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.
The REAlease Kits consist of the respective fluorochrome-conjugated REAlease Complexes and the REAlease Support Kit for removal of the REAlease Complexes and optional relabeling with different fluorochrome-conjugated REAlease Complexes.

Detailed product information

Technical specifications

CloneREAL269
Clonalitymonoclonal
Isotype controlControl Antibody
Hostcell line
Type of antibodyReleasable antibodies, Primary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD279 (PD1)
Distribution of antigenT cells, B cells, myeloid cells
RRIDAB_2751275, AB_2784106, AB_2784105, AB_2751447

Resources for CD279 (PD1) Antibody, anti-mouse,
REAlease
®

References for CD279 (PD1) Antibody, anti-mouse,
REAlease
®

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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®

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