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BUV615 Rat Anti-Mouse CD279 (PD-1)
BUV615 Rat Anti-Mouse CD279 (PD-1)
Flow cytometric analysis of CD279 (PD-1) expression on resting and activated viable Mouse splenic leucocytes. C57BL/6 Mouse splenocytes were either not activated (Left Plot) or activated (Right Plot) by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 567114/567115/553057) 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 BD Horizon™ BUV615 Rat IgG2a, κ Isotype Control (Cat. No. 613005; dashed line histograms) or BD Horizon™ BUV615 Rat Anti-Mouse CD279 (PD-1) antibody (Cat. No. 568601/568602; solid line histograms) at 1.0 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence 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) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 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 viable Mouse splenic leucocytes. C57BL/6 Mouse splenocytes were either not activated (Left Plot) or activated (Right Plot) by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 567114/567115/553057) 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 BD Horizon™ BUV615 Rat IgG2a, κ Isotype Control (Cat. No. 613005; dashed line histograms) or BD Horizon™ BUV615 Rat Anti-Mouse CD279 (PD-1) antibody (Cat. No. 568601/568602; solid line histograms) at 1.0 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence 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) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
製品詳細
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BD Horizon™
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.

推奨アッセイ手順

   BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® 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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

   For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Ultraviolet 615 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. CF™ is a trademark of Biotium, Inc.
抗体の詳細
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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.

フォーマットの詳細
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BUV615
The BD Horizon Brilliant™ Ultraviolet 615 (BUV615) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. BUV615, driven by BD innovation, is designed to be excited by the ultraviolet laser (355 nm) and detected using an optical filter centered near 615-nm (e.g, 610/20 bandpass filter). The acceptor dye can be excited by the Blue (488-nm) and yellow-green (561-nm) lasers resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV615
Ultraviolet 355 nm
350 nm
615 nm
引用&参考文献
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Development References (5)

  1. Bu MT, Yuan L, Klee AN, Freeman GJ. A Comparison of Murine PD-1 and PD-L1 Monoclonal Antibodies.. Monoclon Antib Immunodiagn Immunother. 2022; 41(4):202-209. (Clone-specific: Blocking, Flow cytometry). View Reference
  2. 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
  3. 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
  4. 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
  5. Puzey MS, Craig CJ. Hydrometrocolpos--a case report.. S Afr Med J. 1992; 81(6):336-7. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.