Application
Western blot
Sample
Human Cell lysate - whole cell (Colorectal cancer HCT116)
Loading amount
10 µg
Specification
Colorectal cancer HCT116
Gel Running Conditions
Non-reduced Denaturing (10% Gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/3000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C
Secondary antibody
Name
Non-Abcam antibody was used: Anti-rabbit
Host species: Goat
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL
Exposure
30 second(s)
Bands
Specific: About 55 kDa Non-specific: 34, 43, 72, 95, 130 kDa
Additional data
Abcam response
This customer received a discount code to test this antibody in human. Their preliminary results suggest that this antibody may not be suitable with human samples. Perhaps blocking overnight with 5% BSA instead of milk and using more dilute primary and secondary incubations will improve the non-specific banding. But it is also important to note that the WB should be run in reduced and denatured conditions.
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.
Ms. Injoo Hwang
Verified customer
提交于 Nov 09 2009