Clone:
REA1165
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
PDCD1, PD1, SLEB2, hPD-1, hPD-l, hSLE1

Extended validation for CD279 (PD1) Antibody, anti-human, 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 REA1165
EH12.1++
EH12.2H7++
J105++
MIH4-
NAT105++
PD1.3.1.3++
Cells were incubated with an excess of purified unconjugated CD279 (PD1) (REA1165) 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 CD279 (PD1). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 16 hours 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. 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 CD279 (PD1). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 16 hours 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. 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 CD279 (PD1). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 16 hours 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. 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 CD279 (PD1). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 16 hours 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. 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 CD279 (PD1). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 16 hours 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. 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 CD279 (PD1). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 16 hours 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. 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 CD279 (PD1). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 16 hours 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. 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 CD279 (PD1). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 16 hours 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. 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 CD279 (PD1) (REA1165). 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) (REA1165). 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) (REA1165). 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-human, REAfinity™

Overview

Clone REA1165 recognizes the human CD279 antigen, also known as programmed death-1 (PD-1) or PD1. CD279 is a 55 kDa transmembran protein belonging to the CD28/CTLA-4 family. It is expressed on T cells, B cells, NK cells, activated myeloid cells, and dendritic cells. CD279 is not expressed on naive T cells but induced after activation and expressed on late, differentiated T cells. The ligands PD-L1 (CD274, also knwon as B7-H1) and PD-L2 (CD273, also known as B7-DC) belong to the B7 immunglobulin superfamily. PD-1 and its ligands mediate inhibitory signals that regulate the balance between T cell activation, tolerance, and immunopathology.
Additional information: Clone REA1165 displays negligible binding to Fc receptors.

Alternative names

PDCD1, PD1, SLEB2, hPD-1, hPD-l, hSLE1

Detailed product information

Technical specifications

CloneREA1165
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, non-human primate
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
AntigenCD279 (PD1)
Alternative names of antigenPDCD1, PD1, SLEB2, hPD-1, hPD-l, hSLE1
Molecular mass of antigen [kDa]29
Distribution of antigenB cells, monocytes, T cells, thymocytes, spleen
Entrez Gene ID5133
RRIDAB_2752069, AB_2752075, AB_2752070, AB_2752076, AB_2752071, AB_2752077, AB_2752072, AB_2752078, AB_2752073, AB_2752074

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

Certificates

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

Publications

  1. Okazaki, T. and Honjo, T. (2007) PD-1 and PD-1 ligands: from discovery to clinical application. Int. Immunol. 19: 813-824
  2. Keir, M. E. et al. (2008) PD-1 and its ligands in tolerance and immunity. Annu. Rev. Immunol. 26: 677-704
  3. Serriari, N.-E. et al. (2010) B and T lymphocyte attenuator is highly expressed on CMV-specific T cells during infection and regulates their function. J. Immunol. 185: 3140-3148
  4. Fife, B. T. et al. (2011) The role of the PD-1 pathway in autoimmunity and peripheral tolerance. Ann. N. Y. Acad. Sci. 1217: 45-59

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