Description
The 493 antibody reacts with a cell-surface protein of 130-140 kDa expressed on immature B lymphocytes and a small fraction of newly formed B cells, but not on mature B lymphocytes. The antigen's distribution was defined through the use of antibodies to CD24 (Heat Stable Antigen), IgM, IgD, and CD45R/B220, which are commonly used to discriminate B-cell maturation stages. 493 mAb reacts with the majority of B220+ cells in bone marrow and a fraction of B220+ B cells in spleen, which are CD24[high], IgM[high], and IgD[low]. Cells binding 493 mAb were not detected in thymus, lymph nodes, or peritoneal cavity. This result suggested that 493 mAb does not stain B-1 B cells (CD5+ B lymphocytes), which are particularly found in the peritoneal cavity. The 493 mAb does not seem to have any biological effect when incubated with immature B lymphocytes. It has been observed that the staining pattern of mAb 493 is similar to that of mAb AA4.1, that both antibodies precipitate molecules of the same molecular weight, and that staining by mAb AA4.1 is not blocked by mAb 493, suggesting that the antibodies recognize separate epitopes of the same Early B Lineage antigen.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD Optibuild Brilliant reagents) 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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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 BV650 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. 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.
8. BD Horizon Brilliant Violet 650 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
9. Alexa Fluor® is a registered trademark of Life Technologies Corporation.
10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
