Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblasts)
Loading amount
30 µg
Specification
Mouse Embryonic Fibroblasts
Gel Running Conditions
Reduced Denaturing (10% Bis Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% Milk in PBS w/ .01% tween 20
Secondary antibody
Name
Non-Abcam antibody was used: Goat Anti-Rabbit IgG (Upstate)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
ELC+
Exposure
30 second(s)
Bands
Specific: 20 kDa Non-specific: 45, 50, 60, 150 kDa
Positive control
WT Mouse Embryonic Fibroblasts(MEFs) whole cell lysate
Negative control
None
Additional data
Additional Notes
Very good antibody, with lighter exposures the non specific band are undetectable.
Abcam response
Thank you very much for your feedback. We are very much sure that the non spcecific can be omitted by doing possible protocol optimizations e.g. using range of primary dilutions probably at 1/2000 or 1/3000 dilution and using low conc. of loading lysates e.g. 20ug etc.
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.
Mr. Brian Koss
Verified customer
投稿 Jan 06 2011