User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization

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Project Description/Abstract
“Towards insights into cyclic AMP compartmentalization in cell of lung origin” a cooperation between the department of Molecular Pharmacology (University Groningen, the Netherlands) and the Leibniz-Institut für Molekulare Pharmakologie (Berlin, Germany). These are the entries of a Master internship from February 'till September 2010.

Protocols

 * Taking cells from -80 °C stock
 * Preparation of Medium
 * Preparation Heat inactivated FBS
 * Splitting cells
 * Counting cells
 * Cell stimulation BERLIN
 * Viability staining |Invitrogen protocol
 * HBSS Buffer
 * ELISA (Coating buffer)
 * Co-immunoprecipitation Part I,   Part II &  Part III
 * cAMP-precipitation Part I & Part II
 * RIPA Buffer (Berlin)
 * RIPA Buffer (Groningen)
 * Protein concentration determination: Pierce determination & Bradford assay
 * Coomassie staining
 * Western blot
 * Stripping blot
 * RII Overlay (Radioactivity!)
 * Potter-Elvehjem Cell Homogenizing
 * RNA isolation protocol

Results
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ELISA

 * 1)  Viability &  ELISA
 * 2) Dose effect curve ELISA
 * 3)  ELISA
 * 4)  ELISA
 * 5) ELISA Berlin Okay
 * 1) ELISA Berlin Okay

Co-IP

 * 1) Co-IP 1
 * 2) Co-IP 2
 * 3) Co-IP 3
 * 4) Co-IP 4

RII Overlay

 * 1) RII overlay Okay
 * 1) RII overlay Okay

Misc.

 * Ht31P toxicity
 * Statistical analysis
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 * Presentation [[Image:PRESENTATION_POWERPOINT_August2010.ppt|August]]
 * Presentation [[Image:PRESENTATION_POWERPOINT_August2010.ppt|August]]

Convenient Links

 * Excel2wiki
 * Antibody template

Downloads
   [[Media:BLANCO_ELISA_COPY_ME_BEFORE_USE.xls| Blanc for IL-8 ELISA (Linear regression)]]

   [[Media:BLANCO_PROTEIN_SIZE.xls| Blanc for protein size on gel/membrane (non-Linear regression)]]

   [[Media:BLANCO_IF_COPY_ME_BEFORE_USE.xls.xls| Blanc form IF coverslips]]

Table of contents

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