Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Glioblastoma (GL261))
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
10000 cells
Treatment
HDAC inhibitors for 24hrs
Specification
Mouse Glioblastoma (GL261)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% BSA in 1X TBS with 0.1% Tween-20
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Goat Anti-Rabbit IgG Antibody from Millipore
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL from WestDura (Thermo Fischer)
Exposure
30 second(s)
Additional data
Additional Notes
1. PVDF membrane (Millipore) activated with methanol for 1-2min before preparing the transfer sandwich.
2. Protease inhibitor cocktail was used.
3. For cell lysis Laemmli buffer was used.
4. Bio-Rad wet transfer was performed for 3hrs at 40volts
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Abcam user community
Verified customer
投稿 Sep 12 2014