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Flow cytometric analysis of CD279 (PD-1) expression on resting and activated mouse splenocytes. C57BL/6 mouse splenic leucocytes were either not activated (Left Plot) or activated (Right Plot) by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3 antibody (Cat. No. 553057) for 3 days (37°C). The cells were harvested, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; dotted line histograms) or PE Rat Anti-Mouse CD279 (PD-1) antibody (Cat. No. 568260/568261; solid line histograms) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescent histograms showing the expression of CD279 (PD-1) (or Ig Isotype control staining) were derived from gated events with the forward and side light- scatter characteristics of viable (DAPI-negative) splenocytes. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ PE Rat Anti-Mouse CD279 (PD-1)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and CompBead to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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Companion Products
The 29F.1A12 monoclonal antibody specifically recognizes CD279 which is also known as Programmed Death-1 (PD-1). CD279 (PD-1) is a ~55-kDa type I transmembrane glycoprotein that is encoded by Pdcd1 (Programmed cell death 1) which belongs to the CD28/CTLA-4 family of immunoreceptors within the Ig superfamily. CD279 (PD-1) is comprised of an extracellular region with an IgV-like domain, a transmembrane sequence, and an intracellular region with an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM) that are associated with suppressive immunoregulatory functions. CD279 (PD-1) is variably expressed on some thymocyte subsets and developing B lymphocytes at the pro-B-cell stage. It is also inducibly expressed on activated myeloid cells, B cells, and T cells including exhausted T cells found in mice during chronic viral infections or cancer. Although this co-inhibitory receptor plays roles in mediating immunological tolerance and preventing autoimmune responses it can also inhibit protective immune responses against microbial infections and cancer. CD273 (also known as PD-L2 or B7-DC) and CD274 (PD-L1 or B7-H1) are members of the B7 family within the Ig superfamily. These molecules serve as ligands for CD279 (PD-1) and are variably expressed on lymphoid and nonlymphoid cell types including antigen-presenting cells and tumor cells. The 29F.1A12 antibody can reportedly block the binding of these ligands as well as other mouse PD-1-specific antibodies including clones J43, G4, and RMP1-14. Antibody-mediated inhibition of the interaction between PD-1 and its ligands can serve as an immune checkpoint blockade that can augment T-cell responses against tumor cells.
Development References (4)
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Liang SC, Latchman YE, Buhlmann JE, et al. Regulation of PD-1, PD-L1, and PD-L2 expression during normal and autoimmune responses.. Eur J Immunol. 2003; 33(10):2706-16. (Immunogen: Flow cytometry, Immunofluorescence, Immunohistochemistry). View Reference
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Lázár-Molnár E, Gácser A, Freeman GJ, Almo SC, Nathenson SG, Nosanchuk JD. The PD-1/PD-L costimulatory pathway critically affects host resistance to the pathogenic fungus Histoplasma capsulatum.. Proc Natl Acad Sci U S A. 2008; 105(7):2658-63. (Clone-specific: In vivo exacerbation). View Reference
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Polesso F, Munks MW, Rott KH, Smart S, Hill AB, Moran AE. PD-1-specific "Blocking" antibodies that deplete PD-1+ T cells present an inconvenient variable in preclinical immunotherapy experiments.. Eur J Immunol. 2021; 51(6):1473-1481. (Clone-specific). View Reference
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Puzey MS, Craig CJ. Hydrometrocolpos--a case report.. S Afr Med J. 1992; 81(6):336-7. (Biology). View Reference
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