Western blot abreview for Anti-AKT1 (phospho S473) [EP2109Y] antibody

Good
Abreviews
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Cell lysate - whole cell (A549)
Specification
A549
Treatment
Serum starvation followed by serum addition together with 100ng/ml of EGF for 0 to 3hrs
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% BSA
Dilution
1/1000

Secondary antibody

Name
Non-Abcam antibody was used: goat-anti-rabbit-IgG-HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
ECL
Bands
Specific: 55 kDa
Exposure
2 minute(s) and 0 second(s)

Additional data

Additional Notes
The signal was too weak used at 1:5000 dilution. We got decent signal using 1:1000 dilution. Also signal was very weak when Ab was diluted in milk. 5% BSA is a better choice. No nonspecific bands, background is low, too. So overall, this Ab is good.
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Submitted Mar 21 2014

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