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Purified Rat Anti-Mouse CD185 (CXCR5)
Purified 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. 553088) and Purified Rat Anti-Mouse CD185 (CXCR5) (Cat. No. 551961), followed by Biotin Mouse Anti-Rat IgG2a (Cat. No. 553894) and PE Streptavidin (Cat. No. 554061). 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 Purified Rat IgG2a, κ Isotype Control (Cat. No. 553927) staining.

Flow cytometric analysis of CD185 (CXCR5) expression on mouse splenocytes. Splenocytes were stained with FITC Rat Anti-Mouse CD45R/B220 (Cat. No. 553088) and Purified Rat Anti-Mouse CD185 (CXCR5) (Cat. No. 551961), followed by Biotin Mouse Anti-Rat IgG2a (Cat. No. 553894) and PE Streptavidin (Cat. No. 554061). 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 Purified Rat IgG2a, κ Isotype Control (Cat. No. 553927) 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)
0.5 mg/ml
AB_394302
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

A multistep staining procedure is recommended to amplify immunofluorescent signals for the flow cytometric analysis of mouse CXCR5 expression:

Step 1:        Incubate 1 million cells with 0.1 - 0.5 µg of Purified Rat Anti-Mouse CXCR5 at 4°C for 15 - 20 minutes. Wash cells two times with staining medium containing sodium azide (e.g., Dulbecco's PBS or tissue culture medium [without phenol red and biotin] with 0.09% sodium azide and 2% heat-inactivated FCS or 0.2% BSA).

Step 2:        Incubate the cells with Biotin Mouse Anti-Rat IgG2a (Cat. No. 553894) at 4°C for 20 minutes. Wash cells two times.

Step 3 :        Incubate the cells with ≤ 0.06 µg of PE Streptavidin (Cat. No. 554061) at 4°C for 20 minutes. Wash two times. Resuspend cells in staining medium and analyze stained cells with a FACScan™ Flow Cytometer (Becton Dickinson, San Jose, CA) using appropriate specificity and compensation controls.

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. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  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.
551961 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).

551961 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
551961 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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551961 Rev. 2

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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.