Application
ChIP
Detection step
Real-time PCR
Sample
Human Cell lysate - nuclear (neuroblastoma cells)
Specification
neuroblastoma cells
Negative control
negative control primers for a region surely not to be expressed. The primers were bought from SABioscience (Quiagen) and are named: negative ChIP control primer #1.
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde 1%
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde 1%
Positive control
GAPDH
Other product details
Concentration
0.2 µg/10^6 cells
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TRIS, NaCL, EDTA, TritonX-100
Additional data
Additional Notes
In absolute values the enrichment in the treatment control sample was 3 fold above background, treatment of cells with Forskolin increased that to 15 fold. Samples were analysed by means of quantitative real time PCR with Primers specific for the CRE region containing CRH promoter. The number of biological repeats was just 2, as it is a preliminary experiment.
4 µg of antibody was used for the chromatin of 20 million Be(2)M17 neuroblastoma cells. Only this amount was tested so far, less antibody could be still effective.
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. Ben Jurek
Verified customer
投稿 May 20 2013