Application
Western blot
Sample
Human Cell lysate - whole cell (Y79 retinoblastoma cell line)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Specification
Y79 retinoblastoma cell line
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: TBS with 0.05 % tween20
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Goat anti-rabbit HRP conjugate
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Exposure
3 minute(s) and 0 second(s)
Bands
Specific: about 180 kDa
Additional data
Additional Notes
Human retinoblastoma cell line Y79 gave a specific signal and a few additional non-specific bands. It will do for my application but I don't know what those extra bands are.
lane1:mock
lane2:control shRNA
lane3:shRNA specific for SMCX
Abcam response
Thank you for your valuable feedback. We are pleased to see the antibody has produced a clean band at the expected size for KDM5C / Jarid1C / SMCX antibody. There are a few non specific bands at a lower molecular weight, and we can recommend that with some protocol optimization, for example decreasing the antibody concentrations or blocking in a higher concentration of BSA (5%), this may reduce the amount of non specific binding.
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 Sep 12 2008