Description
The 2B11/CXCR4 monoclonal antibody specifically binds to mouse CD184, which is also known as the C-X-C Chemokine Receptor type 4 , CXCR4. CXCR4 (previously known as Fusin and LESTR), is a seven-transmembrane, G-protein-coupled receptor. It is the specific receptor for the CXC chemokine, SDF-1/CXCL12. Mouse CXCR4 shows 91% homology at the amino acid level with human CXCR4. CXCR4 is widely expressed by hematopoietic and non-hematopoietic cell types including neutrophils, monocytes, T cells, B cells, CD34-positive progenitor cells, endothelial cells, neurons and astrocytes. Human CXCR4 is used by T-tropic HIV-1 as a co-receptor for viral entry. The mouse Cxcr4 gene has been mapped to chromosome 1.
The Purified Rat Anti-Mouse CD184 antibody can be used for the immunofluorscent staining and flow cytometric analysis of mouse leukocytes and cell lines that express CXCR4 (see figure). A multistep staining procedure is recommended to amplify immunofluorescent signals for the flow cytometric analysis of mouse CXCR4 expression: Step 1: Incubate 10e6 cells with 0.5 µg of Purified Rat Anti-Mouse CD184 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 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 by flow cytometry.
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
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 www.bdbiosciences.com/us/s/resources for technical protocols.
