Purifed Rat Anti-Human CXCR5 (CD185)
Clone RF8B2 (RUO)
- Brand BD Pharmingen™
- Alternative Name CD185; Chemokine C-X-C motif receptor 5; BLR1; MDR-15
- Concentration 0.5 mg/ml
- Isotype Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2b, κ
- Reactivity Human (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen Human CXCR5 (CD185)
- Entrez Gene ID 643
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
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Preparation and Storage
Store undiluted at 4°C.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
The Purified Rat Anti-Human CXCR5 (CD185) antibody can be used for the immunofluorescent staining and flow cytometric analyses of human leukocytes and cell lines that express CXCR5 (see Figure). A multistep staining procedure is recommended to amplify immunofluorescent signals for the flow cytometric analysis of human CXCR5 expression:
Step 1: Incubate 10e6 cells with 0.06 - 0.5 mg of purified Purified Rat Anti-Human CXCR5 (CD185) antibody 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 IgG2b (Cat. No. 553898) at 4°C for 20 minutes. Wash cells two times.
Step 3: Incubate the cells with ≤ 0.06 mg of streptavidin-phycoerythrin (Cat. No. 554061) at 4°C for 20 minutes. Wash two times. Resuspend cells in staining medium and analyze stained cells by flow cyotmetry, using appropriate specificity and compensation controls.