Anti-BCRP, clone 5D3, Phycoerythrin conjugated
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | FC | Mouse | Phycoerythrin | Monoclonal Antibody |
Description:
Anti-BCRP, clone 5D3, Phycoerythrin conjugated
Trade Name:
Chemicon (Millipore)
Specificity:
Specifically recognizes an external epitope of the human form of ABCG2. Reportedly reacts with mouse stemcell lines (Ginis, 2004), however mouse reactivity is unsupported at this time.
Immunogen:
3T3 cells expressing human ABCG2
Clone:
5D3
Isotype:
IgG2bκ
Background Information:
Breast cancer resistance protein (BCRP1), also known as placenta-specific ATP-binding cassette (ABC) protein (ABCP) or ABC G-subfamily member 2 (ABCG2), or mitoxantrone resistance protein (MXR), is a member of the ATP-binding cassette superfamily of drug transporters thought to be involved in multi-drug resistance in human neoplastic disease. ABCG2 is expressed in drug-resistant breast, colon and gastric cancer and fibrosarcoma cancer cell lines and in blast cells from patients with acute myeloid leukemia (AML). Normal tissues that express the ABCG2 protein include placental syncytiothrophoblasts, mammary ducts and lobules, epithelium in the large intestine and colon, venous and capillary epithelium and bile canalicular membrane of the liver. Several studies indicate that ABCG2 is also expressed in stem cells of different tissues and may be useful as a stem cell marker. ABCG2 is thought to be responsible for the ability of a population of very primitive hematopoietic and non-hematopoietic cells, the so-called SP (side population) subset, to efflux the Hoechst 33342 fluorescent dye. Semiquantitative RT-PCR results indicate that ABCG2 mRNA is expressed at higher levels in SP cells as compared to non-SP cells in human, rhesus monkey and mouse hematopoietic tissues, with the exception of erythroblasts and NK cells. In human bone marrow, ABCG2 mRNA expression is higher in phenotypically defined populations of primitive CD34+CD38- hematopoietic cells as compared to more differentiated CD34+CD38+ progenitor cells. Bcrp1 mRNA is also expressed in SP cells in murine skeletal muscle, in neural stem cell and progenitor cells and in nestin-positive pancreatic islet derived progenitor cells. Over-expression of the human ABCG2 gene by retroviral-mediated gene transfer confers the SP phenotype to mouse bone marrow cells. Bcrp1 mRNA is expressed in SP as well as non-SP fractions of mouse embryonic stem cells. This suggests that Bcrp1 protein levels may not always be reflected by mRNA levels, or that other characteristics of stem cells, for example, cell cycle status or expression of other transporters, are also involved in the SP phenotype.
Species Reactivity:
Human
Application Notes:
Flow Cytometry 10μL/test
Suggested protocol: Non-adherent cell lines.
1) Pre-Wash the cells 3 times with PBS containing 2% fetal calf serum (FCS) or BSA (fraction V, filter through a 0.45μm membrane).
2) To cell pellet, block cells by adding 10 μL of normal goat serum to the cell pellet mix well gently with pipet tip and incubate for 5 minutes at room temperature (20~25°C).
3) Add 30 μL of the anti-BCRP1/ABCG2 monoclonal antibody (10 μg/mL) diluted with PBS containing 2% FCS or 1% BSA. Mix well, and incubate for 30 minutes at room temperature (20~25°C).
4) Wash by adding 1 mL of PBS containing 2% FCS or 1% BSA, followed by centrifugation at 3,000 rpm for 1 minute at room temperature (20~25°C). Remove supernatant by careful aspiration. Can be repeated once if background is high.
5) Wash by adding 1 mL of PBS containing 2% FCS or 1% BSA followed by centrifugation at 3,000 rpm for 1 minute at room temperature (20~25C), repeat 1X. Remove supernatant by careful as piration.
6) Add 500 μL of PBS containing 2% FCS or 1% BSA and analyze by a flow cytometer.
Optimal working dilutions must be determined by end user.
Suggested protocol: Non-adherent cell lines.
1) Pre-Wash the cells 3 times with PBS containing 2% fetal calf serum (FCS) or BSA (fraction V, filter through a 0.45μm membrane).
2) To cell pellet, block cells by adding 10 μL of normal goat serum to the cell pellet mix well gently with pipet tip and incubate for 5 minutes at room temperature (20~25°C).
3) Add 30 μL of the anti-BCRP1/ABCG2 monoclonal antibody (10 μg/mL) diluted with PBS containing 2% FCS or 1% BSA. Mix well, and incubate for 30 minutes at room temperature (20~25°C).
4) Wash by adding 1 mL of PBS containing 2% FCS or 1% BSA, followed by centrifugation at 3,000 rpm for 1 minute at room temperature (20~25°C). Remove supernatant by careful aspiration. Can be repeated once if background is high.
5) Wash by adding 1 mL of PBS containing 2% FCS or 1% BSA followed by centrifugation at 3,000 rpm for 1 minute at room temperature (20~25C), repeat 1X. Remove supernatant by careful as piration.
6) Add 500 μL of PBS containing 2% FCS or 1% BSA and analyze by a flow cytometer.
Optimal working dilutions must be determined by end user.
Control:
MCF-7 cells
Presentation:
Liquid in PBS, pH 7.4, with 0.1% sodium azide.
Storage Conditions:
Maintain refrigerated at 2-8°C in undiluted aliquots for up to 12 months.Protect from light, do not freeze.
UniProt Number:
Entrez Gene Number:
Gene Symbol:
- ABCG2
- BCRP
- BMDP
- BCRP1
- MXR
- ABC15
- ABCP
- EST157481
- MGC102821
- MXR1
- MRX
- CDw338
Alternate Names:
Breast Cancer Resistance Protein; Placenta-specific ATP Binding Cassette Transporter; ATP-binding Cassette, Sub Family G, Member 2 (ABCG2)
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Key Applications:
Flow Cytometry
Entrez Gene Summary:
The membrane-associated protein encoded by this gene is included in the superfamily of ATP-binding cassette (ABC) transporters. ABC proteins transport various molecules across extra- and intra-cellular membranes. ABC genes are divided into seven distinct subfamilies (ABC1, MDR/TAP, MRP, ALD, OABP, GCN20, White). This protein is a member of the White subfamily. Alternatively referred to as a breast cancer resistance protein, this protein functions as a xenobiotic transporter which may play a major role in multi-drug resistance. It likely serves as a cellular defense mechanism in response to mitoxantrone and anthracycline exposure. Significant expression of this protein has been observed in the placenta, which may suggest a potential role for this molecule in placenta tissue.
UniProt Summary:
FUNCTION: SwissProt: Q9UNQ0 # Xenobiotic transporter that may play an important role in the exclusion of xenobiotics from the brain. May be involved in brain-to-blood efflux. Appears to play a major role in the multidrug resistance phenotype of several cancer cell lines. When overexpressed, the transfected cells become resistant to mitoxantrone, daunorubicin and doxorubicin, display diminished intracellular accumulation of daunorubicin, and manifest an ATP- dependent increase in the efflux of rhodamine 123.
SIZE: 655 amino acids; 72314 Da
SUBUNIT: Monomer or homodimer; disulfide-linked.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Highly expressed in placenta. Low expression in small intestine, liver and colon.
PTM: Glycosylation-deficient ABCG2 is normally expressed and functional.
SIMILARITY: SwissProt: Q9UNQ0 ## Belongs to the ABC transporter family. ABCG (White) subfamily. & Contains 1 ABC transporter domain.
SIZE: 655 amino acids; 72314 Da
SUBUNIT: Monomer or homodimer; disulfide-linked.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Highly expressed in placenta. Low expression in small intestine, liver and colon.
PTM: Glycosylation-deficient ABCG2 is normally expressed and functional.
SIMILARITY: SwissProt: Q9UNQ0 ## Belongs to the ABC transporter family. ABCG (White) subfamily. & Contains 1 ABC transporter domain.
Stem Cell Type:
- Neural Stem Cells
- Hematopoietic Stem Cells
Stem Cell Workflow Stage:
Characterization
Brand Family:
Chemicon
Product Name:
Anti-BCRP, clone 5D3, Phycoerythrin conjugated
Antibody Type:
Monoclonal Antibody
Qty/Pk:
100 assays
Format:
Phycoerythrin
Host:
Mouse