Two-color flow cytometric analysis of CXCR5 (CD185) expression on human peripheral blood lymphocytes. Whole blood was stained with PE Mouse Anti-Human CD19 antibody (Cat. No. 555413/561741) and either BD Horizon™ APC-R700 Rat IgG2b, κ Isotype Control (Cat. No. 564984; Left Plot) or BD Horizon APC-R700 Mouse Anti-Human CXCR5 (CD185) antibody (Cat. No. 565191; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-color flow cytometric contour 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™ LSR II Flow Cytometer System.
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Two-color flow cytometric analysis of CXCR5 (CD185) expression on human peripheral blood lymphocytes. Whole blood was stained with PE Mouse Anti-Human CD19 antibody (Cat. No. 555413/561741) and either BD Horizon™ APC-R700 Rat IgG2b, κ Isotype Control (Cat. No. 564984; Left Plot) or BD Horizon APC-R700 Mouse Anti-Human CXCR5 (CD185) antibody (Cat. No. 565191; Right Plot). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-color flow cytometric contour 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™ LSR II Flow Cytometer System.
Product Details
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_2739103
Aqueous buffered solution containing BSA, protein stabilizer, glycerol 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 APC-R700 under optimum conditions, and unconjugated antibody and free BD Horizon APC-R700 were removed.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
APC-R700 Rat IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
564984
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PE Mouse Anti-Human CD19 RUO
Size
100 Tests
Cat No.
555413
565191 Rev. 2
Antibody Details
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.
This antibody was conjugated to BD Horizon APC-R700, which has been developed exclusively by BD Biosciences as a better alternative to Alexa Fluor® 700. APC-R700 excites and emits at similar wavelengths to Alexa Fluor® 700 yet exhibits significantly improved brightness. This dye can be excited by the red laser and detected with the same filter set as Alexa Fluor® (eg, 730/45-nm filter).
565191 Rev. 2
Format Details
APC-R700
The BD Horizon™ APC-R700 (APC-R700) Dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of an Allophycocyanin (APC) dye donor that has excitation maximum (Ex Max) of 651-nm and an acceptor dye, R700, with an emission maximum (Em Max) at 706-nm. APC-R700, driven by BD innovation, is designed to be excited by the red (627–640-nm) laser and detected using an optical filter centered near 710-nm (e.g., a 720/40-nm bandpass filter). APC-R700 is a brighter alternative to Alexa Fluor™ 700. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC-R700
Red 627-640 nm
651 nm
706 nm
565191 Rev.2
Citations & References
View product citations for antibody "565191" on CiteAb
Development References (6)
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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
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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
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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
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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
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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
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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
565191 Rev. 2
Please refer to Support Documents for Quality Certificates
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.