Description
The 5B11 monoclonal antibody specifically recognizes mouse CD123, the α chain subunit (IL-3Rα) of the mouse IL-3 receptor. The IL-3Rα chain is a 60-70 kD transmembrane glycoprotein that binds IL-3 with low affinity but cannot transduce signals by itself. The mouse IL-3Rα chain can complex with either of two homologous β chain subunits (βc and βIL-3) to form high-affinity heterodimeric IL-3 receptors. The βc chain is the common β chain subunit that can complex with the α subunits of the mouse IL-3R, IL-5R and GM-CSFR to form high-affinity receptors. The βIL-3 subunit can form high affinity IL-3 receptor complexes with only the IL-3Rα subunit. The βIL-3 subunit clone is specific for mouse IL-3 and binds with low affinity. The immunogen used to generate the 5B11 hybridoma was CTLL cells transfected with the mouse IL-3R alpha chain. The 5B11 antibody does not block the binding of IL-3 protein to the high affinity IL-3 receptor heterodimer.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794 or 566349). When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values. For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
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 BB700 under optimal conditions that minimize unconjugated dye and antibody.
1. This antibody was developed for use in flow cytometry.
2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
3. Researchers should determine the optimal concentration of this reagent for their individual applications.
4. An isotype control should be used at the same concentration as the antibody of interest.
5. 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.
6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
9. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
10. Cy is a trademark of GE Healthcare.
