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FACS detection of BCRP1 expression using Murine anti-Human BCRP1 (Catalogue No. MAB4155) monoclonal antibody (red line) or secondary antibody only (bl...
Immunofluorescence Analysis:
Representative image from a previous lot.
Confolcal IF analysis of A431cells using anti-BCRP1 Mouse mAB (Red). ...
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Anti-BCRP1, clone 5D3
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, R | FC, IC, IF, FUNC, WB | Mouse | Purified | Monoclonal Antibody |
Description:
Anti-BCRP1, clone 5D3
Trade Name:
Chemicon (Millipore)
Specificity:
Specifically recognizes an external epitope of the human form of ABCG2. Reportedly reacts with mouse stem cell lines (Ginis, et al., 2004), however mouse reactivity is unsupported at this time.
Molecular Weight:
72 kDa
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
- Rat
Application Notes:
Immunocytochemistry:
Cell cultures fixed with 4% PFA 5-15 minutes at room temperature. Blocking with 10% NGS, 0.1% BSA in PBS with 0.3% Triton X-100 (Ginis et al., 2004).
Flow Cytometry:
1:100-1:1000 dilution from a previous lot was used.
Inhibition Assay:
A previous lot was used to inhibit ABCG2 (Watanabe, K. et al., (2004. FEBS Letters 565:6-10). See also Sarkadi, B., et al., 2004 FEBS 567:116-120,. Sarkadi reports that the detection and inhibition by 5D3 strongly depends on the actual conformation of ABCG2.
Blocking with 10% NGS, 0.1% BSA in PBS with 0.3% triton X-100 (Ginis, 2004).
Optimal working dilutions must be determined by end user.
Cell cultures fixed with 4% PFA 5-15 minutes at room temperature. Blocking with 10% NGS, 0.1% BSA in PBS with 0.3% Triton X-100 (Ginis et al., 2004).
Flow Cytometry:
1:100-1:1000 dilution from a previous lot was used.
Inhibition Assay:
A previous lot was used to inhibit ABCG2 (Watanabe, K. et al., (2004. FEBS Letters 565:6-10). See also Sarkadi, B., et al., 2004 FEBS 567:116-120,. Sarkadi reports that the detection and inhibition by 5D3 strongly depends on the actual conformation of ABCG2.
Blocking with 10% NGS, 0.1% BSA in PBS with 0.3% triton X-100 (Ginis, 2004).
Optimal working dilutions must be determined by end user.
Control:
MCF-7 cells
Quality Assurance:
Routinely evaluated by Western Blot on L6 lysates.
Western Blot Analysis:
1:500 dilution of this lot detected BCRP1 on 10 μg of L6 lysates.
Western Blot Analysis:
1:500 dilution of this lot detected BCRP1 on 10 μg of L6 lysates.
Purification Method:
Protein A purfied
Presentation:
Purified mouse monoclonal IgG2bκ liquid in buffer containing PBS, pH 7.4, with 0.1% sodium azide.
Storage Conditions:
Stable for 1 year at 2-8ºC from date of receipt. Do not freeze.
UniProt Number:
Entrez Gene Number:
Gene Symbol:
- ABCG2
- BCRP
- BMDP
- BCRP1
- MXR
- ABC15
- ABCP
- EST157481
- MGC102821
- MXR1
- MRX
- CDw338
Alternate Names:
- ABC transporter
- ATP-binding cassette transporter G2
- ATP-binding cassette, sub-family G (WHITE), member 2
- ATP-binding cassette, sub-family G, member 2
- Breast cancer resistance protein
- CD338 antigen
- Mitoxantrone resistance-associated protein
- Placenta-specific ATP-binding cassette transporter
- mitoxantrone resistance protein
- placenta specific MDR protein
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
- Immunocytochemistry
- Immunofluorescence
- Affects Function
- Western Blotting
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-BCRP1, clone 5D3
Concentration:
1 .0 mg/mL
Antibody Type:
Monoclonal Antibody
Qty/Pk:
100 µg
Format:
Purified
Host:
Mouse