Application
ChIP
Sample
Arabidopsis thaliana Tissue lysate - nuclear (seedling, 300 mg fresh weight.)
Specification
seedling, 300 mg fresh weight.
Type
Native ChIP (N-ChIP)
Detection step
Real-time PCR
Positive control
I used antibodies that I know work as a positive control for the whole experiment.
qPCR for positive controls used in various publications, e.g. 5' regions of housekeeping genes (actin, At4g04040, both of which are active in my tissue).
qPCR for positive controls used in various publications, e.g. 5' regions of housekeeping genes (actin, At4g04040, both of which are active in my tissue).
Negative control
qPCR for At4g03770 and At4g03880 (silenced transposons)
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C
Additional data
Additional Notes
I was having a hard time getting reproducable results using this antibody. I tried both native and non-native ChIP. For the non-native ChIP, I sonicated my samples until the majority of the DNA was in the range of ~300-800 bp. For native ChIP, I confirmed the digest on a gel.
I used 2-4 ul of antibody.
The negative controls usually came out very clean, but I was having trouble reproducing my positive controls.
This antibody was predicted to react with Arabidopsis, but had not been tested in this species. I would recommend using a different antibody for Arabidpsis work.
code: 100%ABR-T77U0
I used 2-4 ul of antibody.
The negative controls usually came out very clean, but I was having trouble reproducing my positive controls.
This antibody was predicted to react with Arabidopsis, but had not been tested in this species. I would recommend using a different antibody for Arabidpsis work.
code: 100%ABR-T77U0
Abcam response
We appreciate customer's collaboration in testing the antibody in ChIP for Arabidopsis samples.
Consistent sonication conditions are very important for reproducable results (cool samples, no foaming, similar average sheared band size)!
An option would be to use more antibody about 10ug to see if stronger and more consistent results are seen.
Consistent sonication conditions are very important for reproducable results (cool samples, no foaming, similar average sheared band size)!
An option would be to use more antibody about 10ug to see if stronger and more consistent results are seen.
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 Feb 02 2011