Application
Western blot
Sample
Rat Tissue lysate - whole (Whole brain lysate - adult)
Loading amount
40 µg
Specification
Whole brain lysate - adult
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5%
Other product details
Concentration
1 µg/ml
Incubation time
1 hour(s) and 0 minute(s)
Secondary antibody
Name
Non-Abcam antibody was used: goat anti-rabbit IgG HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 95 kDa
Additional data
Additional Notes
In the image below: Lane 1, rat whole brain lysate; Lane 2, mouse whole brain lysate. A second band is seen in mouse, the identity of which is unclear. Concentrations of 1ug/ml, 0.5ug/ml and 0.16ug/ml were used; best results were obtained with 1ug/ml. Exposure time was 10min, but bands started to emerge after 5 mins. Gels higher than 8% acrylamide were tried without success; 8% or lower is recommended. Denaturing of samples at 70C or 95C was not successful. Reducing conditions were used and 20% ethanol was employed for the nitrocellulose membrane transfer. All steps performed at RT.
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. Randal Moldrich
Verified customer
投稿 May 18 2006