User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/08/24: Difference between revisions

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* 370 MBq/mL [γ<sup>32</sup>P]-ATP (18.5 MBq/500 µCi Hartmann Analytic)
* 370 MBq/mL [γ<sup>32</sup>P]-ATP (18.5 MBq/500 µCi Hartmann Analytic)
* Blots for RII Overlay
* Blots for RII Overlay
** PKA Co-IP blot [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/29|29July2010]] used for RII overlay [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/03|03August2010]]
** PKA Co-IP blot [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/29|29July2010]] used for RII overlay [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/03|03August2010]] (#8)
** Lysate blot #2 [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/16|16August2010]] (for AKAP18δ-PP)
** Lysate blot #2 [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/16|16August2010]] (for AKAP18δ-PP)(#1)
** Lysate blot [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/13|13August2010]] (for AKAP18δ-L314E)
** Lysate blot [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/13|13August2010]] (for AKAP18δ-L314E)(#2)
** cAMP precipitation blot (2x) [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/23|23August2010]] (for AKAP18δ-L314E & AKAP18δ-PP)
** cAMP precipitation blot (2x) [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/23|23August2010]] (for AKAP18δ-L314E & AKAP18δ-PP)(#11(blot1)/#13(blot2))
** Lysate blot (3 fragments) [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/18|18August2010]] used for immunoblot [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/19|19August2010]] and [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/21|21August2010]]
** Lysate blot (3 fragments) [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/18|18August2010]] used for immunoblot [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/19|19August2010]] and [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/21|21August2010]](#18)
** cAMP precipitation blot (3 fragments) [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/10|10August2010]] used for blot [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/12|12August2010]] & [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/14|14August2010]]
** cAMP precipitation blot (3 fragments) [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/10|10August2010]] used for blot [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/12|12August2010]] & [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/14|14August2010]](#15)


===Method===
===Method===

Revision as of 08:42, 24 August 2010

<html><style type="text/css"> .todo { color: red } .done { color: green} </style></html>

RII Overlay, Pre-IgG 2nd AB, CSE on HEK293, ASMC to S0, Split cells w. Mileh <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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Summary

  • L314E, 10 μM

Materials & Methods

Materials

Method

Western blot Rabbit pre-immune IgG

  • Secondary AB α-Rabbit (1:10.000), 1h @ RT

RII Overlay

CSE stimulation HEK293

  • HEK293 put to S0 yesterday were stimulated with 15% CSE or only S0 (17.30)

Put ASMC to S0

  • Airway smooth muscle cells D9 P24 were put to S0 for Carolines experiment

Notes

RII overlay

  • PKA Co-IP blot was blocked in the isotope lab
  • Not all membranes were activated again, while they were maintained under moist conditions
  • Column had fallen dry and cracked, this was not noticed until after addition of sample. Separation appeared still succesful

Results

  • Rabbit Pre-immune Western blot

Discussion

  • DONE Ow..


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