Application
Western blot
Sample
Human Cell lysate - whole cell (MOLT4)
Gel Running Conditions
Reduced Denaturing (4-20% gradient)
Loading amount
100000 cells
Treatment
Bortezomib 0-20hours
Specification
MOLT4
Blocking step
BSA 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
Dilution
1/10000
Name
Non-Abcam antibody was used: CST Anti-rabbit IgG, HRP-linked Antibody #7074
Host species: Goat
Clonality: Monoclonal
Conjugation: HRP polymer
Host species: Goat
Clonality: Monoclonal
Conjugation: HRP polymer
Detection
Detection method
ECL
Exposure
30 second(s)
Bands
Specific: 38 kDa Non-specific: 70, 90 kDa
Positive control
MOLT4 samples
Additional data
Additional Notes
While main band for cleaved Caspase-12 proform is clearly visible at 38kda, there are several non-specific bands.
Abcam response
Customer has been contacted with troubleshooting advise. The results can be improved by applying variations in antibody dilution factor and blocking buffer and incubation timing.
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.
Abcam user community
Verified customer
提交于 Jun 19 2015