*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.
Description
The 5D3 monoclonal antibody reacts with the extracellular portion of the human ABCG2 protein, also known as Bcrp1 and MXR. The ABCG2 gene, a member of the multi-drug resistance (MDR) family, is highly expressed on primitive ‘side-population’ (SP) stem cells, which are defined by the efflux of fluorescent dyes such as Rhodamine 123 and Hoechest 33342. In the bone marrow, about 0.05% of cells display the low fluorescence and are highly enriched for repopulating cells. The SP cells, which express low or undetectable levels of CD34, have been identified in multiple species. In addition, expression of ABCG2 appears to be highly conserved.
Applications Reported
For research use only, not for diagnostic or therapeutic use. The 5D3 antibody has been reported for use in flow cytometric analysis.
Applications Tested
The 5D3 antibody has been tested by flow cytometric analysis of human ABCG2 transfected cells and peripheral blood leukocytes. This can be used at less than or equal to 0.5 μg per 100 μl blood (or per 1 million cells in 100 μl total staining volume). It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Kim M, Tunquist H et al. 2002. The multidrug resistance transporter ABCG2 (breast cancer resistance protein 1) effluxes Hoechst 33342 and is overexpressed in hematopoietic stem cells. Clin Cancer Res: 8(1):22-8. Scharenber CW, Harkey MA. et al. 2002. The ABCG2 transporter is an efficient Hoechst 33342 efflux pump and is preferentially expressed by immature human hemtopoietic progenitors. Blood 15;99(2):507-12. Zhou, S., J. D. Schuetz, et al. 2001. The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. Nat Med 7(9): 1028-34. Goodell MA, Rosenzweig M, et al. 1997. Dye efflux studies suggest that hematopoietic stem cells expressing low or undetectable levels of CD34 antigen exist in multiple species. Nat Med 3(12):1337-45.
