Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
30 µg
Specification
HeLa
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 23°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS, 0,1% Tween-20
Dilution
1/1000
Secondary antibody
Dilution
1/10000
Name
Non-Abcam antibody was used: Peroxidase goat anti rabbit IgG-antibody PI-1000 V
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Exposure
20 second(s)
Bands
Specific: ~100 kDa Non-specific: 25, 40 kDa
Additional data
Additional Notes
The specific band is not very strong compared to the unspecific bands. With longer exposure times more unspecific bands appear. If you know where to expect the specific band cutting out this part of the blot increases specificity.
In general this antibody works much better on cells than on WB.
Abcam response
Thank you so much for trying this antibody in human samples.
We appreciate very much your efforts, but are sorry to hear that the antibody did not give satisfactory results. The datasheet indicates that the antibody does not react with human samples. In your case, it seems to be reacting but with background bands. We would welcome additional feedback from other customers as well.
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 Aug 24 2012