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APC-R700 Rat Anti-Mouse CD279 (PD-1)
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Consider BD Horizon™ Red 718 Reagents, bright small molecule alternative to APC-R700 with lower spread into the APC detector. More Info #
APC-R700 Rat Anti-Mouse CD279 (PD-1)

Flow cytometric analysis of CD279 (PD-1) expression on resting and activated mouse splenic leucocytes. Splenic leucocytes from a C57BL/6 mouse were cultured without (Left Plot) or with (Right Plot) 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 harvested, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and stained with either BD Horizon™ APC-R700 Rat IgG2b, κ Isotype Control (Cat. No. 564984; dashed line histograms) or BD Horizon APC-R700 Rat Anti-Mouse CD279 (PD-1) antibody (Cat. No. 566876; solid line histograms) at 1 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 CD276 (PD-1) 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™ 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 splenic leucocytes. Splenic leucocytes from a C57BL/6 mouse were cultured without (Left Plot) or with (Right Plot) 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 harvested, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and stained with either BD Horizon™ APC-R700 Rat IgG2b, κ Isotype Control (Cat. No. 564984; dashed line histograms) or BD Horizon APC-R700 Rat Anti-Mouse CD279 (PD-1) antibody (Cat. No. 566876; solid line histograms) at 1 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 CD276 (PD-1) 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™ and FlowJo® software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Horizon™
PD-1; mPD-1; Pdc1; Pdcd1; Ly101
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
Mouse PD-1 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
18566
AB_2869928
Aqueous buffered solution containing protein stabilizer and ≤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.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566876 Rev. 1
Antibody Details
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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.

This antibody was conjugated to BD Horizon APC-R700, which has been developed exclusively by BD Biosciences as a better alternative to Alexa Fluor® 700. APC-R700 excites and emits at similar wavelengths to Alexa Fluor® 700 yet exhibits significantly improved brightness. This dye can be excited by the red laser and detected with the same filter set as Alexa Fluor® (eg, 730/45-nm filter).

566876 Rev. 1
Format Details
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APC-R700
The BD Horizon™ APC-R700 (APC-R700) Dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of an Allophycocyanin (APC) dye donor that has excitation maximum (Ex Max) of 651-nm and an acceptor dye, R700, with an emission maximum (Em Max) at 706-nm. APC-R700, driven by BD innovation, is designed to be excited by the red (627–640-nm) laser and detected using an optical filter centered near 710-nm (e.g., a 720/40-nm bandpass filter). APC-R700 is a brighter alternative to Alexa Fluor™ 700. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
APC-R700
Red 627-640 nm
651 nm
706 nm
566876 Rev.1
Citations & References
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Development References (7)

  1. 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). View Reference
  2. 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
  3. 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
  4. 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
  5. 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: Flow cytometry, Functional assay). View Reference
  6. 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
  7. 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) View Less
566876 Rev. 1

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