Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (hEK293)
Total protein in input
300 µg
Immuno-precipitation step
Other - protein G dynabeads
Specification
hEK293
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: AFC high salt
Western Blot antibody
Western Blot antibody
None used
Additional data
Additional Notes
This immunoprecipitation was followed by mass spectrometry identification of the immunoprecipitated peptides. The same immunoprecipitation protocol was carried out with an unrelated antibody control. After removal of background contaminants, the proteins were sorted based on the number of identified peptides (spectral counts) highest to lowest. The table shows the first few proteins with the highest number of spectral counts identified (after removal of background contaminants). If the intended antigen is at the top of the list, the antibody is judged to be very good. If the intended antigen is lower on the list but proteins above are known components of the antigen's complex, the antibody is also judged to be very good.
Even though this antibody recognizes its intended target, it also immunoprecipitates a lot of other proteins with higher spectral counts, presumably indicating that those are more abundant in the immunoprecipitate. This indicates that the antibody is likely not very specific.
The number of spectral counts provided in this table is not normalized to the protein size. Although the size of the protein analyzed will influence the number of spectral counts recovered, it only makes large difference for very large proteins (over 200 kd) or very small (15 kd), for most others the differences are small and very rarely affect placement of a protein among other co-immunoprecipitated proteins. DBF4 is 76 kD and the placement would be the same with or without normalization.
Abcam response
Thank you so much for testing this antibody in an untested application (IP).
We appreciate your feedback but are sorry to hear the antibody did not work in IP-Mass-spec. We would welcome additional feedback from other customers as well.
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.
DR. Edyta Marcon
Verified customer
投稿 Jun 27 2015