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BV421 Rat Anti-Mouse CD265 (RANK)
BV421 Rat Anti-Mouse CD265 (RANK)

Flow cytometric analysis of CD265 (RANK) expression on RAW264.7 cells. Cells from the RAW 264.7 (Macrophage; ATCC TIB-71) cell line were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; dashed line histogram) or BD Horizon™ BV421 Rat Anti-Mouse CD265 (RANK) antibody (Cat. No. 566727; solid line histogram) at 1 µg/test. The fluorescence histogram showing CD265 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Flow cytometric analysis of CD265 (RANK) expression on RAW264.7 cells. Cells from the RAW 264.7 (Macrophage; ATCC TIB-71) cell line were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; dashed line histogram) or BD Horizon™ BV421 Rat Anti-Mouse CD265 (RANK) antibody (Cat. No. 566727; solid line histogram) at 1 µg/test. The fluorescence histogram showing CD265 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Horizon™
RANK, TNFSF11A, TRANCER, EOF, FEO, OFE, ODFR, PDB2
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a
Mouse RANK Transfected Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2869831
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 two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet for 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  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.
566727 Rev. 1
Antibody Details
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R12-31

The R12-31 monoclonal antibody specifically recognizes Receptor activator of NF-KB (RANK) which is also known as CD265, TNF-related activation-induced cytokine receptor (TRANCE Receptor/TRANCE-R), Osteoclast differentiation factor receptor (ODFR), or Ly109. CD265 (RANK) is a type I transmembrane glycoprotein that is encoded by Tnfrsf11a (Tumor necrosis factor receptor superfamily member 11a). CD265 (RANK) is expressed by cells in various tissues including osteoclasts and dendritic cells. CD265 (RANK) serves as the signaling receptor for RANKL (CD254/TRANCE) and regulates the activation and differentiation of osteoclasts. Signaling through the RANK:RANKL interaction also increases the survival of dendritic cells and their antigen presentation function to T cells.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes.  With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421  can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421  conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

566727 Rev. 1
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
566727 Rev.1
Citations & References
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Development References (3)

  1. Habbeddine M, Verthuy C, Rastoin O, et al. Receptor Activator of NF-kappaB Orchestrates Activation of Antiviral Memory CD8 T Cells in the Spleen Marginal Zone. Cell Rep. 2017; 21(9):2515-2527. (Clone-specific: Flow cytometry). View Reference
  2. Hakozaki A, Yoda M, Tohmonda T, et al. Receptor activator of NF-kappaB (RANK) ligand induces ectodomain shedding of RANK in murine RAW264.7 macrophages. J Immunol. 2010; 184(5):2442-2448. (Clone-specific: Flow cytometry). View Reference
  3. Kamijo S, Nakajima A, Ikeda K et al. Amelioration of bone loss in collagen-induced arthritis by neutralizing anti-RANKL monoclonal antibody. Biochem Biophys Res Commun. 2006; 347(1):124-132. (Immunogen: Flow cytometry). View Reference
566727 Rev. 1

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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.