User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/02/25: Difference between revisions

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(Autocreate 2010/02/25 Entry for User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization)
 
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{|{{table}} width="800"
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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;">Co-immunoprecipitation part III</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>}}
|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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==Summary==
==Summary==
* Short summary
The samples have been incubated with the (PKA) antibody coated beads and today the beads are washed and stored in Laemli buffer
 
==Materials & Methods==
==Materials & Methods==
===Materials===
===Materials===
*  
* Laemli buffer
* [[PBS]]
* syringe
 


===Method===
===Method===
{| border="1"
* Centrifuge sample incubated with beads, 5 min. 14.000 RPM
|+'''6-wells'''
* Remove supernatant<sup>*</sup> (w. syringe)
!#
* Add 1 mL PBS
!<u>1</u>
* Centrifuge, 5 min. 14.000 RPM
!<u>2</u>
* Remove supernatant<sup>*</sup>
!<u>3</u>
* Add 75 μL Laemli buffer
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* Store samples @ -20 °C
!A
** 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
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|+'''24-wells'''
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|+'''96-wells plate'''
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==Results==
==Results==
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==Conclusion==
==Conclusion==
*{{done}} Ow..
*{{done}} Ow..
==Related topics==
* [[SDS_sample_buffer]]
* [[Sauer:bis-Tris_SDS-PAGE,_the_very_best]]
* [[Odom:Western_Blot]]
* [[Carrico:Recipes]]
* [[Griffitts:Common_buffers]]
* [[Griffin:Antibody_Related_Solutions_&_Recipes]]




Revision as of 02:26, 25 February 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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Summary

The samples have been incubated with the (PKA) antibody coated beads and today the beads are washed and stored in Laemli buffer

Materials & Methods

Materials

  • Laemli buffer
  • PBS
  • syringe


Method

  • Centrifuge sample incubated with beads, 5 min. 14.000 RPM
  • Remove supernatant* (w. syringe)
  • Add 1 mL PBS
  • Centrifuge, 5 min. 14.000 RPM
  • Remove supernatant*
  • Add 75 μL Laemli buffer
  • Store samples @ -20 °C
    • Alternatively go directly on to Western blot (Boil samples and spin beads down, bring supernatant onto gel)


* Be careful not to touch beads

Results

  • WHAT?

Conclusion

  • DONE Ow..

Related topics



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