User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/03/11: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Stop stimulation</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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===Method===
===Method===
{| border="1"
* SDS PAGE was prepared as described [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/01| 01March2010]]
|+'''6-wells'''
* Co-IP samples of [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/05| 05March2010]] & [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/25| 25Feb2010]] were put to gel
!#
** 40 μL sample for 05March2010 samples
!<u>1</u>
** 30 μL sample for 25Feb2010 samples
!<u>2</u>
** 15 μL marker
!<u>3</u>
==Notes==
|-
* SDS PAGE failed after 3 samples of 25Feb2010 & all 8 samples of 05March2010 were put to gel, due to leakage of samples that mixed them
!A
** probable cause is stacking gel that hadn't solidified sufficient, perhaps due to APS 10% that had been prepared earlier that day by someone else and that hadn't been stored on ice. However APS is preservable for 1 week @ -20 °C, so half a day @ RT should be possible.
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==Results==
==Results==
*{{todo| WHAT?}}
*GAPDH--> failure, picture Anouk?
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/16#Attachment| Viability staining]]


==Conclusion==
==Conclusion==
*{{done}} Ow..
*{{done}} Ow..
===Related entries===
====Co-IP====
*
====Run 3: Ht31/S<sup>0</sup>====
*[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/05|03March2010 Counting cells D9 (P21)/D12 (P25)]]
*[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/09|09March2010 Putting to S<sup>0</sup> D9 (P21)]]
*[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/10|10March2010 Stimulating cells D9 (P21)]]
*[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/11|11March2010 Ending stimulation & Viability D9 (P21)]]




Revision as of 02:53, 23 March 2010

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

Stop stimulation <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

  • Add secondary ab (anti-mouse) for GAPDH repeat HT4 blot
  • Prepare 2 SDS gels for Co-IP
  • Stop stimulation

Materials & Methods

Materials

Method

  • SDS PAGE was prepared as described 01March2010
  • Co-IP samples of 05March2010 & 25Feb2010 were put to gel
    • 40 μL sample for 05March2010 samples
    • 30 μL sample for 25Feb2010 samples
    • 15 μL marker

Notes

  • SDS PAGE failed after 3 samples of 25Feb2010 & all 8 samples of 05March2010 were put to gel, due to leakage of samples that mixed them
    • probable cause is stacking gel that hadn't solidified sufficient, perhaps due to APS 10% that had been prepared earlier that day by someone else and that hadn't been stored on ice. However APS is preservable for 1 week @ -20 °C, so half a day @ RT should be possible.

Results

Conclusion

  • DONE Ow..

Related entries

Co-IP

Run 3: Ht31/S0



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