Description
The 5D3/CD338 monoclonal antibody specifically binds to an epitope of ABCG2 (BCRP1), a multi-drug resistance protein that is a member of ATP binding cassette (ABC) transporters. It is highly expressed on primitive stem cells as identified by the "side-population" (SP) phenotype. This SP phenotype is based on the efflux of fluorescent dyes such as Rhodamine 123 and Hoechst 33342. The expression of ABCG2 appears to be highly conserved as it has been identified in various species. Studies show that highly purified murine stem cells express BCRP1 mRNA and this expression declines sharply as the stem cells express CD34. The highest levels of BCRP1 mRNA expression have been seen in KDR+ human stem cells. ABCG2/BCRP1 was clustered as CD338 in the HLDA8 workshop.
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads 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).
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. 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.
7. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
