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