Cell-Cell Interactions: Methods and Protocols (Methods in by Sean P. Colgan

By Sean P. Colgan

A flexible choice of with no trouble reproducible cell-cell interplay assays for uncovering mobile interactions on the molecular point, either in vitro and in vivo. The protocols conceal a various set of cell-cell interplay versions in either common and pathological states, are comfortably adaptable to almost any mobile variety and organ approach, and contain fundamental facts and final result research. moreover, the protocols stick to the profitable tools in Molecular Biology™ sequence layout, every one providing step by step laboratory directions, an advent outlining the foundations at the back of the process, lists of the required apparatus and reagents, and pointers on troubleshooting and heading off recognized pitfalls.

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By Sean P. Colgan

A flexible choice of with no trouble reproducible cell-cell interplay assays for uncovering mobile interactions on the molecular point, either in vitro and in vivo. The protocols conceal a various set of cell-cell interplay versions in either common and pathological states, are comfortably adaptable to almost any mobile variety and organ approach, and contain fundamental facts and final result research. moreover, the protocols stick to the profitable tools in Molecular Biology™ sequence layout, every one providing step by step laboratory directions, an advent outlining the foundations at the back of the process, lists of the required apparatus and reagents, and pointers on troubleshooting and heading off recognized pitfalls.

Show description

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Primary antibody dilution buffer: 1% (w/v) BSA in PBS-T20. 6. Primary antibody: Rabbit anti-ZO-1 (Zymed, South San Francisco, CA). 7. Secondary antibody: Pork anti-rabbit IgG-HRP (Dako Cytomation, Hamburg, Germany). 8. ECL Western blotting detection reagent and Hyperfilm ECL (Amersham Bioscience). 5. Immunofluorescence 1. Eight-well chamber slides (Permanox; Nunc, Roskilde, Denmark). 2. 2% gelatin for 36 to 48 h at 37°C. 3. The methanol used to fix cells should be ice cold. 4. ) and rabbit polyclonal antibody against ZO-1 (Zymed).

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Robey, R. , Nadjem, T. , et al. (2003) Mutations at amino-acid 482 in the ABCG2 gene affect substrate and antagonist specificity. Br. J. Cancer 89, 1971–1978. 36. Cornwell, M. , Safa, A. , Felsted, R. , Gottesman, M. , and Pastan, I. (1986) Membrane vesicles from multidrug-resistant human cancer cells contain a specific 150- to 170-kDa protein detected by photoaffinity labeling. Proc. Natl. Acad. Sci. USA 83, 3847–3850. 37. Rabindran, S. , Collins, K. , and Greenberger, L. M. (1998) Reversal of a novel multidrug resistance mechanism in human colon carcinoma cells by fumitremorgin C.

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