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.
Notes
Interesting entries
Protocols
Results
Convenient Links
Downloads
Blanc for IL-8 ELISA (Linear regression)
Blanc for protein size on gel/membrane (non-Linear regression)
Table of contents
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| Groningen
| Berlin
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- 10/02/2010: Splitting ASM cells
- 12/02/2010: Counting cells
- 15/02/2010: Putting cells to S0
- 16/02/2010: Stimulation hTERT cells
- 17/02/2010: Viability staining
- 18/02/2010: ELISA
- 19/02/2010: Counting and splitting cells
- 22/02/2010: Preparing new DMEM S+
- 23/02/2010: Putting cells to S0, Co-immunoprecipitation
- 24/02/2010: Co-immunoprecipitation part II, (Stimulation hTERT cells v2)
- 25/02/2010: Co-immunoprecipitation part III & Viability staining
- 26/02/2010: ELISA, counting/splitting cells
- 01/03/2010: Western blot
- 03/03/2010: Co-IP day I, GAPDH, VASP, Stop Stimulation HT4
- 04/03/2010: Co-IP day II, ELISA HT4, Cell Stimulation
- 05/03/2010: Co-IP day III, Stop stimulation hTERT, Split hTERT
- 08/03/2010: Western Blot
- 09/03/2010: hTERT stimulation @ splitting, Western blot contn'd
- 10/03/2010: Start stimulation D9, Stop stimulation D12
- 11/03/2010: Stop stimulation
- 12/03/2010: Splitting & Counting cells
- 15/03/2010: Co-IP & ELISA prep
- 16/03/2010: ELISA & hTERT stimulation
- 17/03/2010: Ht31P toxicity
- 18/03/2010: Statistical analysis
- 22/03/2010: Co-IP & Western blot
- 23/03/2010: Co-IP Part II, hTERT Stimulation & Western blot cont'd
- 24/03/2010: Co-IP Part III, Stimulation stop/Viability staining
- 25/03/2010: ELISA
- 26/03/2010: Last Groninger protocols, cell splitting
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- 31/03/2010: Building smoke machine, cell splitting
- 01/04/2010: Creating RII overlay protocol
- 06/04/2010: Putting cells to S0 and splitting cells
- 07/04/2010: Preparation first Pilot Berlin
- 08/04/2010: cAMP precipitation for RII overlay & New cells
- 09/04/2010: Western blot cAMP precipitation for RII overlay
- 11/04/2010: Splitting D9
- 13/04/2010: RII overlay, splitting cells
- 14/04/2010: RII overlay & Western blot
- 15/04/2010: SDS-PAGE (MARKERS) and Immunoblot
- 16/04/2010: Split cells for ELISA and secondary antibody Immunoblot
- 18/04/2010: Refreshed medium for ELISA cells
- 19/04/2010: Put cells to S0 for cAMP precipitation, looked into viability staining (MTT Assay)
- 20/04/2010: Put cells to S0 for ELISA, stimulation for cAMP precipitation
- 21/04/2010: cAMP precipitation, stimulation for ELISA
- 22/04/2010: Stopping ELISA stimulation, preparing gels
- 23/04/2010: ELISA
- 26/04/2010: SDS-PAGE & Refreshed medium
- 27/04/2010: RII Overlay
- 28/04/2010: RII Overlay & Split cells
- 29/04/2010: SDS-PAGE& Finalizing RII overlay
- 30/04/2010: Splitting cells, Immunoblot
- 03/05/2010: Split cells for RII overlay
- 05/05/2010: Put cells to S0 for RII overlay
- 06/05/2010: Stimulation for RII overlay & Preparation of HBBS buffer for Alamar blue
- 07/05/2010: Potter-Elvehjem Cell Homogenizing
- 09/05/2010: SDS-PAGE of cAMP precipitation
- 10/05/2010: Refreshed medium
- 11/05/2010:Split cells
- 13/05/2010:Split cells for ELISA
- 16/05/2010:Put cells to S0 for ELISA
- 17/05/2010:Cell stimulation for ELISA
- 18/05/2010:RII Overlay & Stop stimulation ELISA
- 19/05/2010: RII overlay cAMP precipitation
- 25/05/2010: Splitting cells
- 26/05/2010: New cells, cAMP-precipitation, AKAP79 immunoblot of cAMP-precipitation
- 27/05/2010: cAMP precipitation & AKAP79 immunodetection
- 28/05/2010: Immunoblot AKAP79 (upstate)
- 31/05/2010: SDS-PAGE for AKAP79 immunoblot & RII overlay (HEK293)
- 01/06/2010: SDS-PAGE for immunoblot (fail)
- 02/06/2010: Preparing TRIS Urea gel
- 03/06/2010: Tris urea gel
- 07/06/2010: Urea-TRIS page, splitting cells
- 08/06/2010: Creating RNA isolation protocol
- 09/06/2010: Refreshed medium
- 10/06/2010: Put cells to S0, split cells
- 11/06/2010: Cell stimulation
- 12/06/2010: Stop stimulation: Protein and RNA/Protein isolation
- 13/06/2010: Putting cells to S0, splitting cells and continuing of protein isolation
- 14/06/2010: Cell stimulation
- 15/06/2010: Stop stimulation: Protein and RNA/Protein isolation (2)
- 16/06/2010: Stimulation of cells
- 17/06/2010: Stop stimulation: Protein and RNA/Protein isolation (3)
- 21/06/2010: New HEK293, Bradford assay, cAMP precipitation
- 22/06/2010: cAMP precipitation
- 23/06/2010: Immunoblot
- 24/06/2010: Immunoblot repeat
- 25/06/2010: Coomassie staining
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