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PE Rat Anti-Mouse CD279 (PD-1)
PE Rat Anti-Mouse CD279 (PD-1)
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.
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.
Product Details
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BD Pharmingen™
PD-1; Pdc1; Pdcd1; mPD-1; programmed cell death 1 protein
Mouse (QC Testing)
Rat IgG2a, κ
PD-1 cDNA and PD-1-Ig Fusion Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

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

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. An isotype control should be used at the same concentration as the antibody of interest.
568260 Rev. 1
Antibody Details
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29F.1A12

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.

568260 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
568260 Rev.1
Citations & References
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Development References (4)

  1. 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
  2. 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
  3. 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
  4. Puzey MS, Craig CJ. Hydrometrocolpos--a case report.. S Afr Med J. 1992; 81(6):336-7. (Biology). View Reference
View All (4) View Less
568260 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.