Application
Western blot
Sample
Human Cell lysate - whole cell (Breast cancer cell-lines)
Loading amount
20 µg
Specification
Breast cancer cell-lines
Gel Running Conditions
Non-reduced Denaturing (4-20% Nupage gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Other product details
Dilution
1/1000
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 22°C · Diluent: 5% Milk in TBS-TWEEN
Secondary antibody
Name
Non-Abcam antibody was used: anti-mouse secondary
Host species: Donkey
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
ECL+
Exposure
30 second(s)
Bands
Specific: 49 kDa Non-specific: many kDa
Positive control
MCF7 overexpressing cell-line
Additional data
Additional Notes
We were trying to detect the endogenous c-myc in breast cancer cell-lines. Although the antibody picks up a correct sized band we detected lots of larger background bands particularly one at 75KDa and one at 115KDa.
Abcam response
We thank the researcher for their honest feedback. We do guarantee both the application and species used for c-Myc antibody [9E11] - ChIP Grade (ab56), hence believe with thorough optimisation successful results can be obtained.
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. Cherie Blenkiron
Verified customer
提交于 Jul 18 2008