Application
Western blot
Sample
Rat Cell lysate - whole cell (Hek 293T)
Loading amount
60 µg
Specification
Hek 293T
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/2000
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: TBS-t
Secondary antibody
Name
Non-Abcam antibody was used: Anti-Rabbit (HRP conjugated)
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+
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 22.8, 20 kDa
Additional data
Additional Notes
Using 5% milk as blocking buffer for 30 minutes does not cause non-specific band. Those bands are verified by another anti-WNK (Q256) antibody. T58A mutant on WNK1 1-220 may cause some conformational change and misfolding. Higher molecular weight (22.8 kD) band is missing.
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 18 2010