Technical Data
B0808-09
Bcrp1 (ABCG2) (APC)
Description:
Hematopoietic stem cells are known to express a membrane transporter molecule, known as P-glycoprotein (Pgp), that is encoded by the multidrug resistance gene 1 (MDR1) (1, 2). Expression of Pgp appears to confer a proliferative advantage to stem cells through its anti-apoptotic effects (3, 4). A recent finding provides evidence that an additional transporter molecule known as Bcrp1 (Breast cancer resistance protein 1) or ABCG2 (ATP-binding cassette gene 2), first identified in a breast cancer cell line (5), is expressed on stem cells (6). Bcrp1/ABCG2 belongs to a family of molecules that span the cell membrane six times and can exist as either homo or hetero dimers linked by a short intracellular flexible linker region that
plays an important role in the efflux of a wide range of substrates (7). Although these transporter molecules have initially been thought to play a role in drug resistance, lately they have been found to have utility in better characterizing primitive stem cells. For example, the “side-population” of hematopoietic stem cells, characterized by their inability to retain high levels of the intracellular staining dyes Hoechst 33342 and Rhodamine 123, has been found to express high levels of Bcrp1/ABCG2. Of interest is the observation that Bcrp1/ABCG2 function has been linked to the efflux of the Hoechst dye (6). Furthermore, there is now evidence that this monoclonal can be used as a cell surface marker to identify hematopoietic stem
cells within the bone marrow fraction of lineage negative cells (6). The expression of Bcrp1/ABCG2 appears greatest on CD34--cells and is downregulated with the acquisition of CD34 on the cell surface (6). The ATP binding cassette transporter gene literature and how it impacts stem cell research has been recently reviewed by K. Bunting (8).

Applications:
Suitable for use in Flow Cytometry. Other applications not tested.

Recommended Dilution:
Flow Cytometry: Designed to quantitatively determine the percentage of cells bearing 4-1BB Ligand/TNFSF9 within a population and qualitatively determine the density of 4-1BB Ligand/TNFSF9 on cell surfaces by flow cytometry. Washed cells are incubated with the phycoerythrin-labeled monoclonal antibody, which binds to cells expressing 4-1BB Ligand/TNFSF9. Unbound phycoerythrin-conjugated antibody is then washed from the cells. Cells expressing 4-1BB Ligand/TNFSF9 are fluorescently stained, with the intensity of staining directly proportional to the density of expression of 4-1BB Ligand/TNFSF9. Cell surface expression of 4-1BB Ligand/TNFSF9 is determined by flow cytometric analysis using 488nm wavelength laser excitation and monitoring emitted fluorescence with a detector optimized to
collect peak emissions at 565-605nm.

Storage and Stability:
May be stored at 4°C. For long term storage, aliquot and store at 4C. Do not freeze. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer. Note: We do not recommend adding glycerol as a cryopreservative.
TypeIsotypeCloneGrade
MabIgG2b9H68Purified
SizeStorageShippingSourceHost
100Tests4°C Do not freezeBlue IceHumanMouse
Concentration:
Not Determined
Immunogen:
Synthetic peptide corresponding to human ABCG2.
Purity:
Purified
Form
Supplied as a liquid in saline, 0.5% BSA, 0.1% sodium azide. Labeled with APC.
Specificity:
Recognizes human ABCG2.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Rocchi, E. et al., (2000) Biochem. Biophys. Res. Commun. 271:42. 2. Zhou, S. et al., (2001) Nat. Med. 7:1028.