User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/02/23: Difference between revisions

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(Autocreate 2010/02/23 Entry for User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization)
 
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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;">Putting cells to S<sup>0</sup>, Co-immunoprecipitation</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
Cells were put to S<sup>0</sup> as was supposed to be done [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/22| yesterday]]. [http://en.wikipedia.org/wiki/Immunoprecipitation Co-immunoprecipitation] was performed
 
==Materials & Methods==
==Materials & Methods==
===Materials===
===Materials===
*  
* DMEM S<sup>0</sup>
* DMEM S<sup>+</sup>
* Cells prepared [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/19| 19Feb2010]]
* [[PBS]]
* HT4 cells (neuroblastoma cell line)


===Method===
===Method===
====Putting cells to S<sup>0</sup>====
*Cells were put to S<sup>0</sup> as described [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/15| 15Feb2010]]
** Cells were washed three times with PBS
====Splitting HT4 cells====
* Split HT4 cells as described [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/10| for hTERT cells]]
** Cells of three flasks were resuspended in total volume of 18 mL (3 times 1 mL Trypsin + 15 mL DMEM S<sup>+</sup>)
** 3 mL of suspension was diluted in 42 mL of DMEM S<sup>+</sup>
** Cells were plated out 15 mL per flask
====Co-immunoprecipitation====
*
{| border="1"
{| border="1"
|+'''6-wells'''
|+'''6-wells'''
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==Conclusion==
==Conclusion==
*{{done}} Ow..
*{{done}} Ow..
==Related topics==
* [[Griffin:_Ultimate_Immunoprecipitation_Guide]]




Revision as of 05:21, 23 February 2010

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

Putting cells to S0, Co-immunoprecipitation <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

Cells were put to S0 as was supposed to be done yesterday. Co-immunoprecipitation was performed

Materials & Methods

Materials

  • DMEM S0
  • DMEM S+
  • Cells prepared 19Feb2010
  • PBS
  • HT4 cells (neuroblastoma cell line)

Method

Putting cells to S0

  • Cells were put to S0 as described 15Feb2010
    • Cells were washed three times with PBS

Splitting HT4 cells

  • Split HT4 cells as described for hTERT cells
    • Cells of three flasks were resuspended in total volume of 18 mL (3 times 1 mL Trypsin + 15 mL DMEM S+)
    • 3 mL of suspension was diluted in 42 mL of DMEM S+
    • Cells were plated out 15 mL per flask

Co-immunoprecipitation

6-wells
1 2 3
A
B
24-wells
1 2 3 4 5 6
A
B
C
D
96-wells plate
01 02 03 04 05 06 07 08 09 10 11 12
A
B
C
D
E
F
G
H
I

Results

  • WHAT?

Conclusion

  • DONE Ow..

Related topics



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