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Anti-ABCG2 [5D3]

Ab02927-10.0
Absolute Antibody
ApplicationsFlow Cytometry, ImmunoFluorescence, ImmunoHistoChemistry
Product group Antibodies
TargetABCG2
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Overview

  • Supplier
    Absolute Antibody
  • Product Name
    Anti-ABCG2 [5D3]
  • Delivery Days Customer
    7
  • Application Supplier Note
    This antibody is recommended for identification of human ACBG2 in a sample. This antibody was used in the immunostaining of bone marrow cells to establish a direct link between the SP phenotype and ABCG2 expression (PMID: 11533706). The IgG2b version of this antibody was used to identify AML cell lines that express ABCG2 protein using flow cytometry. This antibody inhibits ABCG2 function, presumably by binding to an external epitope and preventing conformational changes in the transporter, and therefore provides a specific inhibitor assay for ABCG2 function (PMID: 12393637). The SP (side population phenotype) analysis of limbal epithelial cells using 5D3 and TPS-A showed that both 5D3 and TPS-A effectively inhibit Hoechst 33342 dye efflux from these cells (PMID: 15135043). Conformational changes in the ABCG2 multidrug transporter result in the modification of its interaction with 5D3 antibody. This antibody therefore cannot be used to identify human ABCG2 in an immunoblot/western blot assay. The binding of this antibody was maximum in the presence of ATP, ADP, or Ko143, whereas the non-hydrolysable ATP analog, adenosine 5-(beta, gamma-imido)triphosphate (AMP-PNP), and nucleotide trapping by vanadate decreased antibody binding. In membranes expressing the ABCG2-K86M mutant, ATP, ADP, and AMP-PNP decreased, whereas Ko143 increased the binding of this antibody. 5D3 can be used as a sensitive tool to reveal intramolecular changes, reflecting ATP binding, the formation of a catalytic intermediate, or substrate inhibition within the transport cycle of the ABCG2 protein (PMID: 15557326). This antibody was also used in a study involving localization of ABCG2 mitoxantrone resistance-associated protein in normal tissues using immunohistochemistry (PMID: 15990223). This antibody was also used in determining ABCG2 expression using immunofluorescent staining (PMID: 15625123).
  • Applications
    Flow Cytometry, ImmunoFluorescence, ImmunoHistoChemistry
  • Applications Supplier
    FC; IHC; IF
  • Certification
    Research Use Only
  • Clonality
    Monoclonal
  • Clone ID
    5D3
  • Gene ID9429
  • Target name
    ABCG2
  • Target description
    ATP binding cassette subfamily G member 2 (Junior blood group)
  • Target synonyms
    ABC transporter; ABC15; ABCP; ATP-binding cassette transporter G2; ATP-binding cassette, sub-family G (WHITE), member 2 (Junior blood group); BCRP; BCRP1; BMDP; breast cancer resistance protein; broad substrate specificity ATP-binding cassette transporter ABCG2; CD338; CDw338; EST157481; GOUT1; mitoxantrone resistance-associated protein; MRX; multi drug resistance efflux transport ATP-binding cassette sub-family G (WHITE) member 2; MXR; MXR1; MXR-1; placenta specific MDR protein; placenta-specific ATP-binding cassette transporter; UAQTL1; urate exporter
  • Host
    Human
  • Isotype
    IgG1
  • Protein IDQ9UNQ0
  • Protein Name
    ATP-binding cassette sub-family G member 2
  • Scientific Description
    This chimeric human antibody was made using the variable domain sequences of the original Mouse IgG2b format for improved compatibility with existing reagents assays and techniques.
  • Reactivity Supplier
    Human
  • Reactivity Supplier Note
    The original antibody was generated by immunizing mice with intact mouse 3T3 fibroblasts expressing the human ABCG2 protein.
  • Storage Instruction
    -20°C,2°C to 8°C
  • UNSPSC
    12352203

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Figure 1. Western blot analysis of BCRP/ABCG2 using anti-BCRP/ABCG2 antibody (A00457-2). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human A549 whole cell lysates, Lane 2: human MDA-MB-453 whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human Caco-2 whole cell lysates, Lane 5: human PC-3 whole cell lysates, Lane 6: rat liver tissue lysates, Lane 7: rat kidney tissue lysates, Lane 8: mouse HEPA1-6 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-BCRP/ABCG2 antigen affinity purified polyclonal antibody (Catalog # A00457-2) at 0.5 microg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for BCRP/ABCG2 at approximately 65-80KD. The expected band size for BCRP/ABCG2 is at 72KD.
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