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BV480 Rat Anti-Human CXCR5 (CD185)
BV480 Rat Anti-Human CXCR5 (CD185)

Two-color flow cytometric analysis of CXCR5 (CD185) expression on human peripheral blood lymphocytes. Whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and either BD Horizon™ BV480 Rat IgG2b, κ Isotype Control (Cat. No. 565649; Left Plot) or BD Horizon BV480 Mouse Anti-Human CXCR5 (CD185) antibody (Cat. No. 566142/566191; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric dot plots showing the correlated expression of CXCR5 (CD185) [or Ig isotype control staining] versus CD19 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Two-color flow cytometric analysis of CXCR5 (CD185) expression on human peripheral blood lymphocytes. Whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and either BD Horizon™ BV480 Rat IgG2b, κ Isotype Control (Cat. No. 565649; Left Plot) or BD Horizon BV480 Mouse Anti-Human CXCR5 (CD185) antibody (Cat. No. 566142/566191; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric dot plots showing the correlated expression of CXCR5 (CD185) [or Ig isotype control staining] versus CD19 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

Product Details
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BD Horizon™
CD185; Chemokine C-X-C motif receptor 5; BLR1; MDR-15
Human (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2b, κ
Human CXCR5 (CD185)
Flow cytometry (Routinely Tested)
5 µl
643
AB_2739540
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 BV480 under optimum conditions, and unconjugated antibody and free BD Horizon BV480 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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  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. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  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 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.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Antibody Details
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RF8B2

The RF8B2 monoclonal antibody specifically binds to the human CXC chemokine receptor, CXCR5. CXCR5 (also known as CD185, BLR-1 NLR and MDR15), a seven transmembrane, G-protein-coupled receptor, is the specific receptor for CXC chemokine, CXCL13/BLC/BCA-1. In peripheral blood, CXCR5 expression is restricted to B lymphocytes and a small subset of CD4+ and CD8+ lymphocytes. The restricted expression pattern of CXCR5 on B cells and follicular T helper cells (Tfh) suggests that this receptor functions as a regulator of B and T cell migration. Stimulation of T cells with anti-CD3 monoclonal antibody leads to the down-regulation of CXCR5.

The antibody was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set.  BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.

Format Details
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BV480
The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BV480
Violet 405 nm
440 nm
479 nm
Citations & References
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Development References (6)

  1. Barella L, Loetscher M, Tobler A, Baggiolini M, Moser B. Sequence variation of a novel heptahelical leucocyte receptor through alternative transcript formation. Biochem J. 1995; 309(3):773-779. (Biology). View Reference
  2. Dobner T, Wolf I, Emrich T, Lipp M. Differentiation-specific expression of a novel G protein-coupled receptor from Burkitt's lymphoma. Eur J Immunol. 1992; 22(11):2795-2799. (Biology). View Reference
  3. Forster R, Emrich T, Kremmer E, Lipp M. Expression of the G-protein--coupled receptor BLR1 defines mature, recirculating B cells and a subset of T-helper memory cells. Blood. 1994; 84(3):830-840. (Immunogen: Flow cytometry, Immunohistochemistry). View Reference
  4. Gunn MD, Ngo VN, Ansel KM, Ekland EH, Cyster JG, Williams LT. A B-cell-homing chemokine made in lymphoid follicles activates Burkitt's lymphoma receptor-1. Nature. 1998; 391(6669):799-803. (Biology). View Reference
  5. Kouba M, Vanetti M, Wang X, Schafer M, Hollt V. Cloning of a novel putative G-protein-coupled receptor (NLR) which is expressed in neuronal and lymphatic tissue. FEBS Lett. 1993; 321(2-3):173-178. (Biology). View Reference
  6. Legler DF, Loetscher M, Roos RS, Clark-Lewis I, Baggiolini M, Moser B. B cell-attracting chemokine 1, a human CXC chemokine expressed in lymphoid tissues, selectively attracts B lymphocytes via BLR1/CXCR5. J Exp Med. 1998; 187(4):655-660. (Biology). View Reference
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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.