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-APE1 antibody [13B8E5C2] - ChIP Grade
Dilution
1/2000
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 - 40%, FT - 40%, IP - 100%.
The absolute values of APE1 in each well (measured using Odyssey scanning system) were as follows: Input - 115, FT - 98, IP - 34 which means that the effiency of IP was about 15-20% only.
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 - 40%, FT - 40%, IP - 100%.
The absolute values of APE1 in each well (measured using Odyssey scanning system) were as follows: Input - 115, FT - 98, IP - 34 which means that the effiency of IP was about 15-20% only.
Abcam response
We are sorry to hear that the antibody did not give optimal results in IP.
An option would be to include 0.1% SDS in the IP dilution buffer or exclude it if not added.
An option would be to include 0.1% SDS in the IP dilution buffer or exclude it if not added.
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.
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投稿 Mar 07 2011