Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (12% polyacrylamide)
Sample
Human Cell lysate - whole cell (Huh7.5 hepatoma cell line)
Specification
Huh7.5 hepatoma cell line
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS + Tween 0,2% + Milk 5%
Dilution
1/20000
Secondary antibody
Name
Non-Abcam antibody was used: Goat Anti-Mouse IgG (H+L)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL Luminata Classico (Millipore)
Bands
Specific: 48 kDa
Exposure
1 minute(s) and 0 second(s)
Negative control
Lane 1 : no cell lysate
Positive control
Different types of lysates were loaded,
Lane 3 : wild-type Huh7.5 cells
Lane 4 and lane 5 : Huh7.5 expressing irrelevant and relevant shRNA, respectively
Additional data
Additional Notes
Lane 1 : no cell lysate
Lane 2 : size markers (top down : 80, 60, 50, 40 30 and 25 kDa)
Lane 3 : Huh7.5 cell lysate
Lane 4 : Huh7.5 expressing irrelevant shRNA cell lysate
Lane 5 : Huh7.5 expressing relevant shRNA cell lysate
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.
Anthony Morel
Verified customer
提交于 Nov 10 2014