Kerr Lab, Nottingham,

ABC Transporter Research

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Here are our lab protocols. They are all MS-Word files. Don't forget that the protocol (a.k.a. Standard Operating Procedure) is not the same as the Risk Assessment, and so I take no responsibility for safety if you download and use these. They are tried and tested in our lab but may not fit all requirements; I'm always happy to advise other researchers on experimental protocols if I can so please get in touch. Sequence analysis video Time course analysis video Agarose gel electrophoresis ATPase assay Calcium phosphate transfection of mammalian cells Chemically competent bacterial cells and transformation Cytotoxicity assay for mammalian cells Densitometry analysis Electro-competent bacterial cells and transformation Gradient centrifugation techniques · iodixanol gradients · sucrose gradients Immunofluorescence Inclusion body preparation Insect cell culture general routines · handling insect cells · baculovirus generation · plaque assays Mammalian cell culture general routines · Stable cell line generation Membrane preparation from tissue culture cells PCR reaction PCR screening of recombinant plasmids Plasmid manipulations Plasmid preparation Polyethyleneimine mediated mammalian cell transfection Poly-lysine coating of tissue culture plates Precipitations of protein by TCA or acetone Protease inhibitors Protein Assays · DC-Brad (Bio-rad) Lowry assay · bicinchonic acid (BCA) assay SDS PAGE gel staining techniques SDS polyacrylamide gel electrophoresis Small and large scale protein expression Western blotting optimising western blots

Highlight Box February 2020. New paper on using FCS to measure ABCG2:drug interaction January 2020. New paper on the constant contact interface in ABCG2. August 2019 new paper with Beth Coyle on the role of ABCB1 in ependymoma. August 2018 new review on the SNPs of ABCG2 June 2018 new paper on drug binding sites in ABCG2. June 2018 collaborative paper with Bishopp group on phosphate response in roots.  Feb 2017 new paper with the Callaghan group locating drug binding sites in ABCB1/P-glycoprotein June 2016 open access review on ABCG2 published in Biochemical Society Transactions Feb 2016 Paper published in Journal of Neuro-oncology on cell migration inhibition in a glioma cell line October 2015: Paper published in BBA Mol Cell Research on ABCG2 dynamics and oligomerisation studied by TIRF microscopy and fluorescence correlation spectroscopy. October 2015. Two new reviews in Biochemical Society Transactions on the history of ABC transporter research, and ABC transporters in brain tumours July 2015: Paper published in Bioscience Reports on mutations in ABCG2 affecting trafficking and drug transport

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