Application
Western blot
Sample
Human Cell lysate - nuclear (Hek293T)
Gel Running Conditions
Reduced Denaturing (4–15% Mini Protean TGX (Bio-Rad))
Loading amount
30 µg
Treatment
Biotin 50 uM for 24 h
Specification
Hek293T
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 3% BSA in PBS-Tween 20 (0.1%)
Dilution
1/5000
Secondary antibody
Dilution
1/4000
Name
Non-Abcam antibody was used: Anti-Chicken-HRP (A9046, sigma)
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Negative control
WT Hek293T not expressing BirA-Fusion protein
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: kDa Non-specific: 120 kDa
Positive control
Hek293T BirA-Fusion protein expressing cells
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.
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投稿 Oct 30 2015