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PE Rat Anti-Mouse CD185 (CXCR5)
PE Rat Anti-Mouse CD185 (CXCR5)
Flow cytometric analysis of CD185 (CXCR5) expression on mouse splenocytes. Splenocytes were stained with FITC Rat Anti-Mouse CD45R/B220 (Cat. No. 551959) and PE Rat Anti-Mouse CD185 (CXCR5) (Cat. No. 551959/561988). Two-color dot plot depicting CD185 (CXCR5) and CD45R/B220 expression were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes. The quadrant markers for the bivariate dot plots were set based on the PE Rat IgG2a, κ Isotype Control (Cat. No. 553930) staining.
Flow cytometric analysis of CD185 (CXCR5) expression on mouse splenocytes. Splenocytes were stained with FITC Rat Anti-Mouse CD45R/B220 (Cat. No. 551959) and PE Rat Anti-Mouse CD185 (CXCR5) (Cat. No. 551959/561988). Two-color dot plot depicting CD185 (CXCR5) and CD45R/B220 expression were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes. The quadrant markers for the bivariate dot plots were set based on the PE Rat IgG2a, κ Isotype Control (Cat. No. 553930) staining.
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_394300
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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Recommended Assay Procedures

The PE-conjugated 2G8 antibody is convenient for the immunofluorescent staining and flow cytometric analyses of mouse leukocytes and cell lines that express CXCR5 (see figure).  Chemokine receptors are known to internalize during manipulation resulting in low frequency expression. Immunophenotyping studies of chemokine receptors need to be performed on freshly collected whole blood (<24 Hrs). Incubation with the antibody should be done in the dark. Cellular manipulation, such as Ficoll separation, freezing, or exposure to cold temperatures prior to staining should be minimized, as these treatments have been shown to cause a decrease in staining intensity and inconsistent results.  

Investigators should note that alternative staining procedures may be necessary.  A multiple-step staining procedure is recommended, in some instances, to amplify immunofluorescent signals for the flow cytometric analysis of mouse 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 FITC Streptavidin (Cat. No. 554060).

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561988 Rev. 2
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).

561988 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
561988 Rev.2
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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561988 Rev. 2

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