User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/08/16: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(7 intermediate revisions by the same user not shown)
Line 5: Line 5:
{|{{table}} width="800"
{|{{table}} width="800"
|-
|-
|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;">SDS-PAGE lysates, Cell splitting</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>}}
|-
|-
Line 26: Line 26:
! align="center" style="background:#f0f0f0;"|<u>10</u>
! align="center" style="background:#f0f0f0;"|<u>10</u>
|-
|-
!Gel<br>cAMP prec.
!Gel<br>Lysates
| align="center"|[[Media:HiMark_protein_marker_-_Invitrogen.jpg|Invitrogen Marker]]<br> (8 μL)
| align="center"|[[Media:HiMark_protein_marker_-_Invitrogen.jpg|Invitrogen Marker]]<br> (8 μL)
| align="center"|Jurkat <br>(20 μL)
| align="center"|Jurkat <br>(20 μL)
| align="center"|HEK293<br>(20 μL)
| align="center"|HEK293<br>(20 μL)
| align="center"|[[Media:Biorad_-_Precision_Plus_Protein_Dual_Color_marker.jpg|Biorad Marker]]<br> (8 μL)
| align="center"|[[Media:Biorad_-_Precision_Plus_Protein_Dual_Color_marker.jpg|Biorad Marker]]<br> (8 μL)
| align="center"|C1<br>(20 μL)
| align="center"|C2<br>(20 μL)
| align="center"|C3<br>(20 μL)
| align="center"|S1<br>(20 μL)
| align="center"|S1<br>(20 μL)
| align="center"|S2<br>(20 μL)
| align="center"|S2<br>(20 μL)
| align="center"|S3<br>(20 μL)
| align="center"|S3<br>(20 μL)     
| align="center"|C1<br>(20 μL)
| align="center"|C2<br>(20 μL)
| align="center"|C3<br>(20 μL)     
|-
|-
|}
|}
Line 329: Line 329:
==Notes==
==Notes==
-->
-->
==Results==
==Results==
*Ponceau staining of blots
*Ponceau staining of blots
[[Image:16082010_Ponceau_Lysates_Blot_-1.TIF|500px]]
[[Image:16082010_Ponceau_Lysates_Blot_-1.TIF|350px]][[Image:Coomassie_gel_after_blot_16.8.10-1.tif|350px]]<br>
[[Image:16082010_Ponceau_Lysates_Blot_-2.TIF|500px]]
[[Image:16082010_Ponceau_Lysates_Blot_-2.TIF|350px]][[Image:Coomassie_gel_after_blot_16.8.10-2.tif|350px]]
 
==Discussion==
==Discussion==
*{{done}} Ow..
*{{done}} Ow..

Revision as of 06:49, 23 August 2010

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

SDS-PAGE lysates, Cell splitting <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

  • 2x SDS-PAGE
4-12% Gradient NuPage Gel loading LYSATES hTERT D9 + HEK293, HeLa -S & Jurkat see codes 21June2010
# 1 2 3 4 5 6 7 8 9 10
Gel
Lysates 		Invitrogen Marker
(8 μL) 		Jurkat
(20 μL) 		HEK293
(20 μL) 		Biorad Marker
(8 μL) 		C1
(20 μL) 		C2
(20 μL) 		C3
(20 μL) 		S1
(20 μL) 		S2
(20 μL) 		S3
(20 μL)
    • NOTE: After 60 min. the SDS-PAGE was put to ice and the voltage was turned from 200 V to 100 V, to latter because of limitation in the amount of people running wet blots.
    • Both gels were blotted using the NuPage wetblot system, 2 h @ 30 V
  • Cells were splitted to D9 P28
  • Cells in 24-wells were put to S0

Results

  • Ponceau staining of blots


Discussion

  • DONE Ow..


Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/16&oldid=447296"