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PerCP-Cy™5.5 Rat Anti-Mouse CD185 (CXCR5)
PerCP-Cy™5.5 Rat Anti-Mouse CD185 (CXCR5)

Flow cytometric analysis of CXCR5 on mouse splenocytes.  Left Panel: Splenocytes from C57BL/6 mice were stained either with a PerCP-Cy™5.5 Rat IgG2a, κ isotype control (Cat. No. 550765; shaded histogram) or with the PerCP-Cy™5.5 Rat Anti-Mouse CXCR5 antibody (Cat. No. 50528; unshaded histogram).  Histograms were derived from gated events based on light scattering characteristics for CD45R/B220+ cells.  Middle and Right Panels: Splenocytes from C57BL/6 mice were stained with both a APC Rat Anti-Mouse CD45R/B220 antibody (Cat.No. 553092) and either a PerCP-Cy™5.5 Rat IgG2a, κ isotype control (middle panel) or the PerCP-Cy™5.5 Rat Anti-Mouse CXCR5 antibody (right panel).  Dot plots were derived from gated events based on light scattering characteristics for splenocytes.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.

Flow cytometric analysis of CXCR5 on mouse splenocytes.  Left Panel: Splenocytes from C57BL/6 mice were stained either with a PerCP-Cy™5.5 Rat IgG2a, κ isotype control (Cat. No. 550765; shaded histogram) or with the PerCP-Cy™5.5 Rat Anti-Mouse CXCR5 antibody (Cat. No. 50528; unshaded histogram).  Histograms were derived from gated events based on light scattering characteristics for CD45R/B220+ cells.  Middle and Right Panels: Splenocytes from C57BL/6 mice were stained with both a APC Rat Anti-Mouse CD45R/B220 antibody (Cat.No. 553092) and either a PerCP-Cy™5.5 Rat IgG2a, κ isotype control (middle panel) or the PerCP-Cy™5.5 Rat Anti-Mouse CXCR5 antibody (right panel).  Dot plots were derived from gated events based on light scattering characteristics for splenocytes.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.

Product Details
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BD Pharmingen™
Blr1; C-X-C chemokine receptor type 5; CXC-R5; CXCR-5; Gpcr6; MDR15
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
Mouse CXCR5
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_1727522
Aqueous buffered solution containing ≤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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

Recommended Assay Procedures

Flow cytometry:  Chemokine receptors are known to internalize during manipulation resulting in low frequency expression.  Investigators are advised to perform immunophenotyping studies of chemokine receptors on freshly collected samples (<24 Hrs). Incubation with the antibody should be done at 4°C in the dark. Cellular manipulation, such as Ficoll separation, freezing, or exposure to cold temperatures prior to staining should be minimized and have been shown to cause a decrease in staining intensity and/or inconsistent results.

Investigators should note that alternative staining procedures may be neccessary.  A multiple-step staining procedure is strongly recommended, in some instances, to amplify immunofluorescent signals for the flow cytometric analysis of mouse CD185 (CXCR5) expression.  Investigators may find the Purified Rat Anti-Mouse CD185 (CXCR5) antibody (Cat. No. 551961) to be useful in conjunction with appropriate secondary and tertiary reagents for detecting low frequency expression, such as with Biotin Mouse Anti-Rat IgG2a (Cat. No. 553894) and PE Streptavidin (Cat. No. 554061) or PerCP-Cy™5.5 Streptavidin (Cat. No. 551419).

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. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  5. This PerCP-conjugated product is sold under license to the following patent: US Patent No. 4,876,190.
  6. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  7. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560528 Rev. 4
Antibody Details
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2G8

The 2G8 monoclonal antibody specifically binds to the mouse C-X-C Chemokine Receptor type 5, CXCR5. CXCR5 is also known as CD185, BLR1, NLR and MDR15. CXCR5 is a seven-transmembrane, G-protein-coupled receptor that is specific for the CXC chemokine, CXCL13/BLC/BCA-1. The expression of CXCR5 has been detected in spleen, lymph nodes, tonsils, brain, bone marrow, T cells, B cells, cerebrum, cerebellum, hippcampus and pituitary. In mouse spleen, CXCR5 was strictly expressed by mature B cells and a small subset of T lymphocytes. CXCR5 plays a role in directing the migration of B and T cells to B cell follicles with the spleen and certain other lymphoid tissues. The immunogen used to generate 2G8 hybridoma was a recombinant protein containing N-terminal amino acids of mouse CXCR5 (GST-NmBLR1).

560528 Rev. 4
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) 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.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
560528 Rev.4
Citations & References
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Development References (7)

  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, Mattis AE, Kremmer E, Wolf E, Brem G, Lipp M. A putative chemokine receptor, BLR1, directs B cell migration to defined lymphoid organs and specific anatomic compartments of the spleen. Cell. 1996; 87(6):1037-1047. (Immunogen). 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. Kaiser E, Forster R, Wolf I, Ebensperger C, Kuehl WM, Lipp M. The G protein-coupled receptor BLR1 is involved in murine B cell differentiation and is also expressed in neuronal tissues. Eur J Immunol. 1993; 23(10):2532-2539. (Biology). View Reference
  6. 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
  7. 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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560528 Rev. 4

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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.