Application
Western blot
Sample
Human Cell lysate - whole cell (MRC5VA Lung Epithelial Fibroblasts)
Loading amount
15 µg
Specification
MRC5VA Lung Epithelial Fibroblasts
Gel Running Conditions
Reduced Denaturing (9%)
Blocking step
Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Other product details
Dilution
1/500
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 23°C · Diluent: 5% Milk in PBS/Tween
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-Rabbit HRP (DAKO)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/1000
Detection
Detection method
ECL+
Exposure
10 minute(s) and 0 second(s)
Positive control
Normal and IR-samples loaded on to the gel.
Additional data
Additional Notes
Unfortunately I ordered the incorrect antibody as this one was not tested for Western Blotting, but I can confirm that it indeed does not work for Western Blotting. Use ab58567 as this does work.
Abcam response
We recommend using BSA as a blocking solution as milk can have damaging effects on phosphorylated tissue. Additionally, it appears the recommended dilutions for the other applications with this antibody are lower than the dilution used in this experiment. It may also help to increase the concentration of the antibody in the future.
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. Darren Arbon
Verified customer
投稿 Apr 14 2011