Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing (4-20% gradient HEPES-glycine gel)
Sample
Human Cell lysate - whole cell (MCF10A cell line)
Specification
MCF10A cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 2% · Temperature: 20°C
Other product details
Incubation time
1 hour(s) and 30 minute(s) · Temperature: 20°C · Diluent: 2% BSA-TBS + 0.1% Tween-20
Dilution
1/1000
Secondary antibody
Name
Non-Abcam antibody was used: anti-rabbit HRP conjugate (PerkinElmer)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
ECL+
Bands
Specific: ~100 kDa
Exposure
15 second(s)
Negative control
siRNA against target protein
Positive control
extract.
-Loading control: anti-eIF2alpha (Abcam: ab26197)
Additional data
Abcam response
We are very pleased that this antibody worked in WB for this customer. However, previous results obtained by the use of this antibody were showing high background and we decided to not recommend it for WB. We are currently in contact with the lab to re-test this antibody in WB in order to confirm its suitability for this application.
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.
Miguel Sanchez
Verified customer
投稿 Mar 27 2014