User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/03/22: Difference between revisions
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m (→Preparing film) |
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* Treat with secondary antibody (diluted in 5% TBST-milk solution) (1h, RT) | * Treat with secondary antibody (diluted in 5% TBST-milk solution) (1h, RT) | ||
* Wash 3x 10 min. in TBST buffer | * Wash 3x 10 min. in TBST buffer | ||
{| border="1" | {| border="1" | ||
|+'''24-wells''' | |+'''24-wells''' | ||
!# | ! align="center" style="background:#f0f0f0;"|# | ||
!<u>1</u> | ! align="center" style="background:#f0f0f0;"|<u>1</u> | ||
!<u>2</u> | ! align="center" style="background:#f0f0f0;"|<u>2</u> | ||
!<u>3</u> | ! align="center" style="background:#f0f0f0;"|<u>3</u> | ||
!<u>4</u> | ! align="center" style="background:#f0f0f0;"|<u>4</u> | ||
!<u>5</u> | ! align="center" style="background:#f0f0f0;"|<u>5</u> | ||
!<u>6</u> | ! align="center" style="background:#f0f0f0;"|<u>6</u> | ||
! align="center" style="background:#f0f0f0;"|<u>7</u> | |||
! | ! align="center" style="background:#f0f0f0;"|<u>8</u> | ||
| | ! align="center" style="background:#f0f0f0;"|<u>9</u> | ||
! align="center" style="background:#f0f0f0;"|<u>10</u> | |||
! align="center" style="background:#f0f0f0;"|<u>11</u> | |||
! align="center" style="background:#f0f0f0;"|<u>12</u> | |||
! align="center" style="background:#f0f0f0;"|<u>13</u> | |||
! align="center" style="background:#f0f0f0;"|<u>14</u> | |||
! align="center" style="background:#f0f0f0;"|<u>15</u> | |||
! | |||
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! | |||
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!11 | |||
!12 | |||
! | |||
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! | |||
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! | |||
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|- | |- | ||
! | !Gel 1 | ||
| | |Marker<br> (15 μL) | ||
| | |D9A 1<br> (all <30 μL) | ||
| | |D9A 2<br> (all <30 μL) | ||
| | |D9A 3<br> (all <30 μL) | ||
| | |D9A 4<br> (all <30 μL) | ||
| | |D12A 1<br> (30 μL) | ||
| | |D12A 2<br> (30 μL ) | ||
| | |D12A 3<br> (30 μL) | ||
| | |D12A 4<br> (30 μL) | ||
| | |D9B 1<br> (30 μL) | ||
| | |D9B 2<br> (30 μL) | ||
| | |D9B 3<br> (30 μL) | ||
|D9B 4<br> (30 μL)|||| | |||
|- | |- | ||
! | ! align="center" style="background:#f0f0f0;"|Gel 2 | ||
| | | align="center" style="background:#f0f0f0;"|Marker<br> (15 μL) | ||
| | | align="center" style="background:#f0f0f0;"|D12B 1<br> (35 μL) | ||
| | | align="center" style="background:#f0f0f0;"|D12B 2<br> (35 μL) | ||
| | | align="center" style="background:#f0f0f0;"|D12B 3<br> (35 μL) | ||
| | | align="center" style="background:#f0f0f0;"|D12B 4<br> (35 μL) | ||
| | | align="center" style="background:#f0f0f0;"|X | ||
| | | align="center" style="background:#f0f0f0;"|D9C 1<br> (35 μL) | ||
| | | align="center" style="background:#f0f0f0;"|D9C 2<br> (35 μL) | ||
| | | align="center" style="background:#f0f0f0;"|D9C 3<br> (35 μL) | ||
| | | align="center" style="background:#f0f0f0;"|D9C 4<br> (35 μL) | ||
| | | align="center" style="background:#f0f0f0;"|D12C 1<br> (35 μL) | ||
| | | align="center" style="background:#f0f0f0;"|D12C 2<br> (35 μL) | ||
| align="center" style="background:#f0f0f0;"|D12C 3<br> (35 μL) | |||
| align="center" style="background:#f0f0f0;"|D12C 4<br> (35 μL) || | |||
|- | |- | ||
|} | |} | ||
# Only beads (30 μL) | |||
# Beads w. antibody (30 μL) | |||
# Beads w. antibody (30 μL) & lysis buffer (200 μL) | |||
# Beads w. antibody (30 μL) & sample (200 μL) | |||
A. [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/23|23Feb2010]] to [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/23|25Feb2010]]<br> | |||
B. [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/03|03March2010]] to [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/05|05March2010]]<br> | |||
C. [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/15|15March2010]] to [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/17|17March2010]] | |||
==Notes== | ==Notes== | ||
* No ice block was put in transfer system for the first 24 min. (of 60 min. in total) | * No ice block was put in transfer system for the first 24 min. (of 60 min. in total) |
Revision as of 07:02, 22 March 2010
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Summary
Materials & MethodsMaterialsMethodCo-immunoprecipitation part I
SDS-PAGE
Western Blot
A. 23Feb2010 to 25Feb2010 Notes
Results
Conclusion
|