The M1/69 monoclonal antibody specifically binds to CD24 (Heat-Stable Antigen, HSA or HsAg), a variably glycosylated, glycosyl-phosphatidylinositol-anchored membrane protein expressed on erythrocytes, granulocytes, monocytes, lymphocytes, and neurons. Hematopoietic stem cells of the embryonic yolk sac and fetal liver express CD24. Levels of expression of CD24 vary during differentiation of the T and B cell lineages. In the bone marrow, hematopoietic progenitors acquire CD24 expression upon commitment to the B-lymphocyte lineage. Immature B cells in the bone marrow express low CD24 levels whereas peripheral B lymphocytes express intermediate to high levels of CD24. The level of CD24 expression has been reported to rise upon activation of splenic B cells with LPS, but not with CD154 (CD40 Ligand). The majority of thymocytes express high levels of CD24, while most mature thymic and peripheral T lymphocytes do not express CD24. In contrast, TCR-bearing thymocytes which emigrate to the spleen are CD24+. Dendritic cells of the thymus, spleen, liver, and epidermal Langerhans cells have also been reported to express CD24. CD24 is not expressed by NK cells, as determined by staining with J11d mAb (Cat. No. 553146). CD24 is involved in the costimulation of CD4+ T cells by B cells, it is a "co-inducer" of in vitro thymocyte maturation, and it is a ligand of CD62P (P-selectin). While the monoclonal antibodies 30-F1, M1/69, and J11d all react with CD24, they show subtle differences in the level of staining of different lymphocyte populations. When possible, investigators should continue to use the same monoclonal anti-CD24 antibody as used in previous studies.
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application. 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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385). Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
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 to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. 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.
2. Researchers should determine the optimal concentration of this reagent for their individual applications.
3. An isotype control should be used at the same concentration as the antibody of interest.
4. 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.
5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. CF™ is a trademark of Biotium, Inc.
10. BD Horizon Brilliant Ultraviolet 615 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
