Application
Western blot
Sample
Human Cell lysate - whole cell (lymphocytes)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
35 µg
Specification
lymphocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk in TBS-T
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: ECL Rabbit IgG, HRP-linked whole Ab
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 280 kDa Non-specific: 185, 80, 65, 48, 25, kDa
Additional data
Additional Notes
This is a 1 minute exposure. RIPA buffer and sigma protease and phosphatase inhibitors used. Lysates also sonicated. 10% MeOH transfer buffer used, nitrocellulose membrane.
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 Jan 30 2014