Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Loading amount
100 µg
Specification
Liver
Gel Running Conditions
Reduced Denaturing (8)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/50
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk
Secondary antibody
Name
Non-Abcam antibody was used: Goat Anti-rabbit IgG, HRP-linked Antibody #7074
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/2000
Detection
Detection method
ECL
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 57 kDa Non-specific: 52, 57, 75, 125 kDa
Positive control
Wild-type Liver lysates from C57BL/6 mice
Negative control
Sesn3-knockout liver lysates from Sesn3[-/-] mice
Additional data
Additional Notes
Also tried 1:500 and 1:100 dillution and used two different blocking methods (milk and Roche WBR) and they gave similar results. Many more non-specific bands were observed if exposed longer. Although SESN3 protein signal is detectable, it is weak and overlayed by nearby non-specific bands.
This experiment was done as a collaboration between Abcam and Lee lab (please check the information below).
This experiment was done as a collaboration between Abcam and Lee lab (please check the information below).
Abcam response
Thank you for your valuable feedback and for participating in our Abreview testing discount program. We are sorry to hear that this antibody has not given you good results in western blot using mouse sample. The concentration could be diluted further (1/500-1/3000) as suggested on the datasheet because one of the cause for non-specific bands is high concentration of antibody. Otherwise, it could that this antibody has not been tested with mouse and is possible that it may not be suitable for use.
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 Nov 11 2011