Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
20 µg
Specification
U2OS cells
Blocking step
(agent) for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Other product details
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: Licor Blocking Buffer
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Alexa Fluor 680 anti-Rabbit antibody
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor 680
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor 680
Detection
Detection method
Licor
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 250 kDa
Additional data
Additional Notes
To check whether this antibody is specific to phosphorylation, I added 1 ul of Alkaline phosphatase (CIP, NEB) and incubated for 30 min at 37. The observed band after phosphatase might be from either short incubation with CIP or non-specific interaction to non- protein.
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 Feb 10 2017