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BV421 Rat Anti-Mouse CD273
BV421 Rat Anti-Mouse CD273
Two-color flow cytometric analysis of CD273 expression on mouse splenic leucocytes (dendritic cells, left and center panels). BALB/c mouse splenic leucocytes were cultured overnight with recombinant mouse GM-CSF (Cat. No. 554586; 5 ng/ml). The cells were harvested and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Hamster Anti-Mouse CD11c antibody (Cat. No. 550261/561119) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or BD Horizon BV421 Rat Anti-Mouse CD273 antibody (Cat. No. 564245; Right Panel). Two-color flow cytometric contour plots show the correlated expression patterns of CD273 (or Ig Isotype control staining) versus CD11c for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Two-color flow cytometric analysis of CD273 expression on mouse splenic leucocytes (dendritic cells, left and center panels). BALB/c mouse splenic leucocytes were cultured overnight with recombinant mouse GM-CSF (Cat. No. 554586; 5 ng/ml). The cells were harvested and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Hamster Anti-Mouse CD11c antibody (Cat. No. 550261/561119) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or BD Horizon BV421 Rat Anti-Mouse CD273 antibody (Cat. No. 564245; Right Panel). Two-color flow cytometric contour plots show the correlated expression patterns of CD273 (or Ig Isotype control staining) versus CD11c for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
Pdcd1lg2; B7dc; B7-DC; PD-1 ligand 2; PD-L2; Btdc
Mouse (Tested in Development)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
Mouse B7-DC
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2738698
Aqueous buffered solution containing BSA 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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

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).

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. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  8. 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.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564245 Rev. 2
Antibody Details
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TY25

The TY25 monoclonal antibody specifically binds to CD273, also known as B7-DC or PD-L2. CD273 is a 42-kDa type I membrane glycoprotein encoded by the Pdcd1lg2 gene of the B7 family of the Ig superfamily. Pdcd1lg2 mRNA is expressed selectively in dendritic cells (DC), liver, and a few transformed cell lines (hepatic, neuroblastoma, and myeloid), but not in lymphoid tissues or macrophages. The protein has not been detected on resting peripheral T and B lymphocytes, macrophages, or DC, but it is upregulated upon activation of T cells, macrophages, and DC by a variety of stimulatory factors. B7-DC's receptor, PD-1, contains an ITIM (Immunoreceptor Tyrosine-based Inhibitory Motif) on its intracytoplasmic region and is expressed on activated B and T lymphocytes, suggesting that B7-DC-PD-1 interaction may be involved in the negative regulation of immune responses. The second PD-1 ligand, B7-H1 (PD-L1), is also a member of the B7 family of the Ig superfamily.  B7-DC may also participate in positive immunoregulation, or costimulation of T cells, through an additional receptor, which is not PD-1 and distinct from the alternate receptor for B7-H1. Moreover, B7-DC is the first B7 ligand that has been found to participate in bi-directional communication; ligation of B7-DC on DC stimulates those DC. The TY25 antibody blocks the binding of B7-DC to PD-1, but not to its alternate ligand.

564245 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
564245 Rev.2
Citations & References
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Development References (11)

  1. Ansari MJ, Salama AD, Chitnis T, et al. The programmed death-1 (PD-1) pathway regulates autoimmune diabetes in nonobese diabetic (NOD) mice. J Exp Med. 2003 July; 198(1):63-69. (Clone-specific: Blocking, Flow cytometry, Immunohistochemistry). View Reference
  2. Carreno BM, Collins M. The B7 family of ligands and its receptors: New pathways for costimulation and inhibition of immune responses. Annu Rev Immunol. 2002; 20:29-53. (Biology). View Reference
  3. Latchman Y, Wood CR, Chernova T, et al. PD-L2 is a second ligand for PD-1 and inhibits T cell activation. Nat Immunol. 2001; 2(3):261-268. (Biology). View Reference
  4. Liu X, Gao JX, Wen J, et al. B7DC/PDL2 promotes tumor immunity by a PD-1-independent mechanism. J Exp Med. 2003; 197:1721-1730. (Clone-specific: Blocking, Flow cytometry). View Reference
  5. Nguyen LT, Radhakrishnan S, Ciric B, et al. Cross-linking the B7 family molecule B7-DC directly activates immune functions of dendritic cells. J Exp Med. 2002; 196:1393-1398. (Biology). View Reference
  6. Salama AD, Chitnis T, Imitola J, et al. Critical role of the programmed death-1 (PD-1) pathway in regulation of experimental autoimmune encephalomyelitis. J Exp Med. 2003 July; 198(1):71-78. (Biology). View Reference
  7. Shin T, Kennedy G, Gorski K et al. Cooperative B7-1/2 (CD80/CD86) and B7-DC costimulation of CD4+ T cells independent of the PD-1 receptor. J Exp Med. 2003; 198:31-38. (Biology). View Reference
  8. Tseng S-Y, Otsuji M, Gorski K, et al. B7-DC, a new dendritic cell molecule with potent costimulatory properties for T cells. J Exp Med. 2001; 193:839-846. (Biology). View Reference
  9. 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. (Clone-specific: Blocking, Immunohistochemistry, In vivo exacerbation). View Reference
  10. Wang S, Bajorath J, Flies DB, Dong H, Honjo T, Chen L. Molecular modeling and functional mapping of B7-H1 and B7-DC uncouple costimulatory function from PD-1 interaction. J Exp Med. 2003; 197:1083-1091. (Biology). View Reference
  11. Yamazaki T, Akiba H, Iwai H, et al. Expression of programmed death 1 ligands by murine T cells and APC. J Immunol. 2002; 169(10):5538-5545. (Immunogen: Flow cytometry, Immunoprecipitation, Western blot). View Reference
View All (11) View Less
564245 Rev. 2

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