User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/02/16: Difference between revisions
m (→Method) |
|||
Line 43: | Line 43: | ||
===Method=== | ===Method=== | ||
* Remove medium | * Remove medium from 24-wells plate with ASMC's grown ON in DMEM (S<sup>0</sup>) | ||
* Rinse twice with warm PBS and remove buffer | * Rinse twice with warm PBS and remove buffer | ||
* Add 200 μL DMEM S<sup>0</sup> with either HT31 or HT31P (see schedule below) | * Add 200 μL DMEM S<sup>0</sup> with either HT31 or HT31P (see schedule below) |
Revision as of 08:22, 16 May 2010
<html><style type="text/css"> .todo { color: red } .done { color: green} </style></html>
Stimulation hTERT cells | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||
SummaryTo look at the effect of Protein kinase A (PKA) and Epac on the IL-8 production by airway smooth muscle cells (ASMC's) induced by cigarette smoke extract (CSE), 8-pcpt-2'-o-me-camp (8-pcpt) and 6-Benz-cAMP (6-Benz) are used to activate Epac and PKA respectively. To illustrate the influence of A kinase Anchoring Proteins (AKAP's) an allround inhibitor of PKA (subunit RII) binding to AKAP's HT31 is used. An inactive peptide HT31P is used as an control. The purpose of the described method is to stimulate the cells with the substances to prepare for future experiments (e.g. ELISA, Western blot) Materials & MethodsMaterials
Stock solutions
Method
CTR = control, CSE = Cigarette Smoke extract, 8-pcpt = 8-pcpt-2'-o-me-camp Related entriesRun 1: Ht31/Ht31P D9/D12 |