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Trouble shooting pouring gels

Trouble shooting pouring gels (see http://www.ruf.rice.edu/~ bioslabs/studies/sds-page/sdsgoofs.html ). Gel caster leaks – Gel doesn’t polmerize - Other – watch out for air bubbles, especially when putting in the combs! -n- butanol or isopropanol on top of separating gel.

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Trouble shooting pouring gels

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  1. Trouble shooting pouring gels (see http://www.ruf.rice.edu/~bioslabs/studies/sds-page/sdsgoofs.html) Gel caster leaks – Gel doesn’t polmerize- Other – watch out for air bubbles, especially when putting in the combs! -n-butanol or isopropanol on top of separating gel

  2. Troubleshooting Western blots Choice of membrane – nitrocellulose vs PVDF Choice of blocking solution – don’t use milk if doing streptavidin-HRP Running conditions – gets hot! Complete transfer. Right direction! High background No bands

  3. Local and distant cell communication

  4. Direct communication

  5. Membrane protein functions

  6. Overview of cell signaling

  7. The structure of a G-protein-linked receptor

  8. The functioning of a G-protein-linked receptor

  9. The structure and function of a tyrosine-kinase receptor

  10. A phosphorylation cascade

  11. http://www.nature.com/icb/journal/v78/n4/images/icb200049f1.gifhttp://www.nature.com/icb/journal/v78/n4/images/icb200049f1.gif

  12. http://www.cellsignal.com/reference/pathway/B_Cell_Antigen.htmlhttp://www.cellsignal.com/reference/pathway/B_Cell_Antigen.html

  13. Far Western Blotting from Pierce.net

  14. Yeast two hybrid http://www.clontech.com/products/detail.asp?tabno=2&catalog_id=630489&page=all

  15. Figure 1. Applications of functional protein microarrays A representative sample of the different assays that have been performed on functional protein microarrays. Proteins are immobilized at high spatial density onto a microscope slide and the slide can then be probed for various interactions. While Cy5 is the fluorophore shown, many other fluorophores can be used for detection http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1828913/pdf/nihms17379.pdf

  16. Figure 4. A. A yeast proteome microarray containing ~4,400 GST-tagged yeast proteins printed in duplicate. The slide was probed with anti-GST antibodies followed by Cy5-labeled anti-rabbit antibodies. 40 of 48 blocks are shown. B. A kinase assay with 33P-γ-ATP and active kinase on a yeast proteome microarray. Dark spots represent radiolabeledphosphorylated substrates on the array. Kinasesthat autophosphorylateare printed in each of the 40 blocks (shown above in blue boxes) and serve as reference points on the slide http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1828913/pdf/nihms17379.pdf

  17. http://www.nature.com/nrm/journal/v4/n1/fig_tab/nrm1007_F1.htmlhttp://www.nature.com/nrm/journal/v4/n1/fig_tab/nrm1007_F1.html

  18. http://qcri.queensu.ca/Greer.html

  19. http://www.plantmethods.com/content/2/1/12

  20. http://130.15.90.245/interest.htm

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