Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Loading amount
50 µg
Specification
U2OS cells
Gel Running Conditions
Reduced Denaturing (4-12%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBST with 1% BSA
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-Rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL
Exposure
15 minute(s) and 0 second(s)
Bands
Specific: 130, 140 kDa Non-specific: 60, 20 kDa
Additional data
Additional Notes
The strongest bands are smaller than expected and might recognize protein fragments resulting from degradation.
Abcam response
Thank you for your valuable feedback. ab99315 seems to recognise specific bands around the expected molecular weight of AZI1 in these experimental conditions. It might be possible to eliminate background bands by using more stringent blocking conditions. Thank you for your time - we hope that this information will be helpful for future customers.
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 Oct 10 2012