Application
Western blot
Sample
Arabidopsis thaliana Tissue lysate - nuclear (50 6-day old Arabidopsis seedlings)
Gel Running Conditions
Reduced Denaturing
Loading amount
0.4 µg
Specification
50 6-day old Arabidopsis seedlings
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk in PBS
Concentration
0.5 µg/ml
Secondary antibody
Dilution
1/10000
Name
Non-Abcam antibody was used: mouse anti-rabbit IgG HRP
Host species: Mouse
Clonality: Polyclonal
Conjugation: HRP polymer
Host species: Mouse
Clonality: Polyclonal
Conjugation: HRP polymer
Detection
Detection method
ECL
Exposure
2 minute(s) and 0 second(s)
Bands
Specific: 17 kDa Non-specific: 17, 20 kDa
Positive control
All three lanes in the image attached are Arabidopsis nuclear extract. The expected size is about 14.3 kDa which wasn't detected.
Additional data
Abcam response
It seems another customer has reported a positive result using Arabidopsis thaliana tissue lysate with ab4174, loading 20ug of sample and using ab4174 at a 1/1000 dilution for a 12H incubation at 4°C diluted in PBST.
It seems that ab4174, does not work under these conditions. Perhaps increasing the amount of protein loaded and the antibody dilution will yield a better result.
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
投稿 Nov 20 2020