Application
Western blot
Loading amount
240000 cells
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Mouse Cell lysate - whole cell (Primary Neural cells)
Specification
Primary Neural cells
Treatment
Supplemented with and without serum in the culture media.
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
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
Dilution
1/5000
Detection
Detection method
ECL from WestDura (Thermo Fischer)
Bands
Specific: 37 kDa Non-specific: many kDa
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.
Abcam response
Our Scientific Support team is currently investigating this negative Abreview, and we will resolve the issue in accordance with our Abpromise.
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
提交于 Sep 16 2014