Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa cells)
Total protein in input
200 µg
Specification
HeLa cells
Immuno-precipitation step
Protein A/G
Other product details
Concentration
5 µg/mg lysate
Incubation time
14 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 50mMTris,pH8.0,50mMKCl,1mMEDTA,5%glycero
Western Blot antibody
Name
Abcam antibody:Anti-DDB1 antibody
Additional data
Additional Notes
Mock IP was performed using 5µg/mg lysate DNA Polymerase delta prebleed serum. IP step was done with ProtA/G for 6h at 4C with shaking using 200 µg of HeLa whole cell extract.
The relative amounts of protein loaded on the gel were as follows: Input - 15%, FT - 15%, IP - 100%.
The absolute values of DDB1 in each well (measured using Odyssey scanning system) were as follows: Input - 95.1, FT - 89.6, IP - 9.3, and mock IP - 8.8 which means that IP did not work at all. And those 10% of DDB1 bound to the beads are actually due to non-specific binding.
The relative amounts of protein loaded on the gel were as follows: Input - 15%, FT - 15%, IP - 100%.
The absolute values of DDB1 in each well (measured using Odyssey scanning system) were as follows: Input - 95.1, FT - 89.6, IP - 9.3, and mock IP - 8.8 which means that IP did not work at all. And those 10% of DDB1 bound to the beads are actually due to non-specific binding.
Abcam response
We are sorry to hear that the customer was unsuccessful in IP.
This is an untested adn therefore not guaranteed application.
This is an untested adn therefore not guaranteed application.
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 Mar 18 2011