Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Gel Running Conditions
Reduced Denaturing (10% Tris-Gly)
Loading amount
30 µg
Treatment
siRNA to CKII alpha prime
Specification
HeLa cells
Blocking step
LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Other product details
Incubation time
3 hour(s) and 0 minute(s) · Diluent: Odyssey® Blocker 1:1 in PBS + 0.1% Tween
Dilution
1/5000
Secondary antibody
Dilution
1/10000
Name
Non-Abcam antibody was used: AlexaFluor 680 goat anti-rabbit
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 680
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 680
Detection
Detection method
Odyssey® system
Negative control
siRNA to CKII alpha prime
Exposure
1 second(s)
Bands
Specific: 39 kDa Non-specific: 30, 36, 53, 59, 63, 67 kDa
Positive control
HeLa whole cell extract
Additional data
Additional Notes
Picture legend
1 - HeLa whole cell extract
2 - siRNA to CKII alpha prime in HeLa cells
All non-specific bands are quite weak.
Abcam response
Thank you for your valuable feedback. As discussed by the author of this Abreview, there are some fainter non specific bands on this blot. However, the specific band is quite intense. We would suggest that the results could be optimized further to eliminate the background bands and still maintain the specific band. For example, try reducing the concentration of antibody.
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.
Dr. Svetlana Khoronenkova
Verified customer
投稿 Jul 22 2010