User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/02/25: Difference between revisions
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* Centrifuge, 5 min. 14.000 RPM | * Centrifuge, 5 min. 14.000 RPM | ||
* Remove supernatant<sup>*</sup> | * Remove supernatant<sup>*</sup> | ||
* Add 75 μL Laemli buffer | * Add 75 μL Laemli buffer to beads | ||
* Store samples @ -20 °C | * Store samples @ -20 °C | ||
** Alternatively go directly on to Western blot (Boil samples and spin beads down, bring supernatant onto gel) | ** Alternatively go directly on to Western blot (Boil samples and spin beads down, bring supernatant onto gel) | ||
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<sup>*</sup> Be careful not to touch beads | <sup>*</sup> Be careful not to touch beads | ||
====[[wikipedia: VASP_(gene)| VASP]]==== | ====[[wikipedia: VASP_(gene)| VASP]]==== | ||
* Use cells grown by Bachelor students {{todo}} | * Use cells grown by Bachelor students {{todo}} | ||
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** Add Buffer B until pH of 9.6 has reached (150 - 200 mL) | ** Add Buffer B until pH of 9.6 has reached (150 - 200 mL) | ||
* Continue as described [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/18#Method| 18Feb2010]] | * Continue as described [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/18#Method| 18Feb2010]] | ||
==Related== | |||
====BSc.==== | |||
* Alamar blue | |||
* Stop stimulation (hTERT) | |||
* Cells to 96-wells (hTERT) | |||
===Results=== | |||
* {{todo|VASP blot, Anouk}} | |||
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/26#Attachment|Viability staining]] | |||
===Related entries=== | |||
====Run 2: Ht31 Dose/Effect Curve & S<sup>0</sup> D9/D12==== | |||
*[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/19|Counting cells 19Feb2010]] | |||
*[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/23|Putting to S<sup>0</sup> 23Feb2010]] | |||
*[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/24|Stimulating cells 24Feb2010]] | |||
==Same action== | |||
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/18| ELISA]] | * [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/18| ELISA]] | ||
==Related topics== | ==Related topics== | ||
* [[SDS_sample_buffer]] | * [[SDS_sample_buffer]] |
Revision as of 02:23, 23 March 2010
<html><style type="text/css"> .todo { color: red } .done { color: green} </style></html>
Co-immunoprecipitation part III | <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> | ||||||||||||
SummaryThe samples have been incubated with the (PKA) antibody coated beads and today the beads are washed and stored in Laemli buffer Materials & MethodsMaterials
MethodCo-immunoprecipitation
VASP
Coating of plate
BSc.
Results
Related entriesRun 2: Ht31 Dose/Effect Curve & S0 D9/D12Same actionRelated topics
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