Application
Western blot
Sample
Human Tissue lysate - whole (Brain)
Loading amount
50 µg
Specification
Brain
Gel Running Conditions
Reduced Denaturing (gel 15 %)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/1000
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: in TBS 1X + milk 1%
Secondary antibody
Name
Non-Abcam antibody was used: GOAT anti-rabbit Ig (H+L) HRP (interchim)
Host species: Goat
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/1000
Detection
Detection method
ECL super signal (Pierce)
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 18, 19 kDa Non-specific: 60, 80 kDa
Positive control
None
Negative control
Brain extract from mouse knockout for alpha-synuclein protein.
Additional data
Additional Notes
We used brain extract from transgenic mouse overexpressing human alpha-synuclein. Brains were homogenized in high salting buffer supplemented with protease and phosphatase coktail inhibitors. We tried both on PVDF and nitrocellulose membranes and the antibody worked in both cases.
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.
Dr. Sébastien Brot
Verified customer
提交于 Jul 05 2011