BUV737 Rat Anti-Mouse CD279 (PD-1)

This product is the replacement for 749306.

BD Horizon™ BUV737 Rat Anti-Mouse CD279 (PD-1)

Name: BUV737 Rat Anti-Mouse CD279 (PD-1)
Brand: BD Horizon™
SKU: 568363

Clone RMP1-30

(RUO)

Flow cytometric analysis of CD279 (PD-1) expression on resting and activated mouse splenocytes. Splenocytes from a C57BL/6 mouse were activated in culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 553057) for 3 days. The resting (Left Plot) or activated (Right Plot) splenocytes were 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™ BUV737 Rat IgG2b, κ Isotype Control (Cat. No. 612762; dashed line histograms) or BD Horizon™ BUV737 Rat Anti-Mouse CD279 (PD-1) antibody (Cat. No. 568363; solid line histograms) at 1.0 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histograms showing CD279 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) splenic 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.

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Product Details

Brand: BD Horizon™

Alternative Name: PD-1; mPD-1; Pdc1; Pdcd1; Ly101

Reactivity: Mouse (QC Testing)

Isotype: Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ

Immunogen: Mouse PD-1 Recombinant Protein

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

Entrez Gene ID: 18566

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: 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 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

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
An isotype control should be used at the same concentration as the antibody of interest.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
BD Horizon Brilliant Ultraviolet 737 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.
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.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer Plus RUO

Size 1000 Tests Cat No. 566385

Brilliant Stain Buffer RUO

Size 1000 Tests Cat No. 566349

BUV737 Rat IgG2b, κ Isotype Control RUO

Size 50 µg Cat No. 612762

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.5 mg Cat No. 553142

View Details

568363 Rev. 1

Antibody Details

RMP1-30

The RMP1-30 monoclonal antibody specifically recognizes CD279 which is also known as PD-1 (programmed death-1). CD279 (PD-1) is a ~55 kDa type I transmembrane glycoprotein that is encoded by Pdcd1 which belongs to the CD28/CTLA-4 family within the Ig superfamily. CD279 (PD-1) is comprised of an extracellular region with an IgV-like domain and an intracellular region with an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM) that are associated with inhibitory signaling functions. CD279 (PD-1) is transiently expressed on CD4-CD8- thymocytes and developing B lymphocytes at the pro-B-cell stage. It is also expressed on activated myeloid cells, B cells, and T cells including exhausted T cells found in mice during chronic viral infections or cancer. This co-inhibitory receptor reportedly functions in negative regulation of immune responses and thus helps guard against autoimmunity and preserves peripheral tolerance. 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 that serve as ligands for CD279 (PD-1). The RMP1-30 antibody reportedly does not block the binding of CD279 (PD-1) to these ligands.

568363 Rev. 1

Format Details

BUV737

The BD Horizon Brilliant™ Ultraviolet 737 (BUV737) 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 735-nm. BUV737, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 740-nm (e.g., 740/35 bandpass filter). The acceptor dye can be excited by the Red (628–640nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BUV737

Excitation Source: Ultraviolet 355 nm

Excitation Max: 350 nm

Emission Max: 735 nm

568363 Rev.1

Citations & References

Development References (7)

Agata Y, Kawasaki A, Nishimura H, et al. Expression of the PD-1 antigen on the surface of stimulated mouse T and B lymphocytes. Int Immunol. 1996 May; 8(5):765-772. (Biology: Flow cytometry). View Reference
Gupta PK, Godec J, Wolski D, et al. CD39 Expression Identifies Terminally Exhausted CD8+ T Cells. PLoS Pathogens. 2015; 11(10):e1005177. (Clone-specific: Flow cytometry). View Reference
Kasagi S, Kawano S, Okazaki T, et al. Anti-programmed cell death 1 antibody reduces CD4+PD-1+ T cells and relieves the lupus-like nephritis of NZB/W F1 mice. J Immunol. 2010; 184(5):2337-2347. (Clone-specific: Flow cytometry). View Reference
Liu X, Gibbons RM, Harrington SM, et al. Endogenous tumor-reactive CD8(+) T cells are differentiated effector cells expressing high levels of CD11a and PD-1 but are unable to control tumor growth. Oncoimmunology. 2013; 2(6):e23972. (Clone-specific: Flow cytometry). View Reference
Matsumoto K, Inoue H, Nakano T, et al. B7-DC regulates asthmatic response by an IFN-gamma-dependent mechanism. J Immunol. 2004; 172(4):2530-2541. (Immunogen). View Reference
Nishimura H, Agata Y, Kawasaki A, et al. Developmentally regulated expression of the PD-1 protein on the surface of double-negative (CD4-CD8-) thymocytes. Int Immunol. 1996 May; 8(5):773-780. (Biology). View Reference
Tsushima F, Iwai H, Otsuki N, et al. Preferential contribution of B7-H1 to programmed death-1-mediated regulation of hapten-specific allergic inflammatory responses. Eur J Immunol. 2003; 33(10):2773-2782. (Biology). View Reference

View All (7)

568363 Rev. 1

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