Summary
- According to the NuPage blotting manual adding 0.01% SDS might improve transfer to the membrane
- Volume of blotting chamber is ~200 mL so adding 100 μL of 20% SDS solution should be fine
- Blot of 16August2010 is repeated (only one gel)
- NOTE: Only 10 µL for HEK293 cells and de Untreated samples a smaller amount of sample left (about 70-80% of CSE treated)
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
(10 μL)
| Biorad Marker
(8 μL)
| C1
(20 μL)
| C2
(20 μL)
| C3
(20 μL)
| S1
(<20 μL)
| S2
(<20 μL)
| S3
(<20 μL)
|
- This gel will be blotted ON @, 30 V using the NuPage system while on ice in the cold room (4°C)
- Blots blocked yesterday were treated with α-RIIα, α-AKAP95 (mouse) and α-AKAP450 to see what we can find in lysates despite the fact that transfer was not complete
Results
- Ponceau staining of membrane and Gel after 15h blotting
 
- Ponceau of the second membrane
Discussion
- Transfer looks still incomplete, and hardly anything seems to have leaked through the first membrane
|
ON Blot lysate samples
Main project page
Previous entry Next entry
