Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Neuroblastoma (Neuro2A))
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
400000 cells
Treatment
Serum deprivation (0.1%) for 24 hrs
Specification
Mouse Neuroblastoma (Neuro2A)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% BSA in 1X TBS with 0.1%
Dilution
1/500
Secondary antibody
Dilution
1/10000
Name
Non-Abcam antibody was used: Goat Anti-Mouse IgG Antibody from Millipore
Host species: Mouse
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Mouse
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL from WestDura (Thermo Fischer)
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 200 kDa Non-specific: No kDa
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.
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提交于 Mar 25 2015