Application
ChIP
Sample
Mouse Cell lysate - whole cell (teratocarcinoma F9 cells)
Specification
teratocarcinoma F9 cells
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Specification of the cross-linking agent: DSG and formaldehyde
Duration of cross-linking step: 0 second(s)
Specification of the cross-linking agent: DSG and formaldehyde
Detection step
Real-time PCR
Positive control
H3K4 trimethyl
Negative control
Rabbit IgG
Other product details
Concentration
0.25 µg/µg chromatin
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: RIPA
Additional data
Additional Notes
Response to additional questions:
IPs of four cell lines were done in parallel, using H3K4me3 as a positive control for succesful IP procedure.
The cells were cross-linked for 1 hour at room temperature with Disuccinimidyl glutarate (DSG, 2mM final conc in PBS),
then rinsed with PBS and cross-linked 10 minutes at room temperature with formaldehyde (1% final concentration).
IPs of four cell lines were done in parallel, using H3K4me3 as a positive control for succesful IP procedure.
The cells were cross-linked for 1 hour at room temperature with Disuccinimidyl glutarate (DSG, 2mM final conc in PBS),
then rinsed with PBS and cross-linked 10 minutes at room temperature with formaldehyde (1% final concentration).
Abcam response
We are greatful for customer's feedback. We have not had similar negative feedback for ab2621.
The problem seems to be with the efficiency of the IP.
This antibody is batch tested in ChIP.
We would recommend the customer to try IP only with formaldehyde fixation for 10min as we do in-house.
The problem seems to be with the efficiency of the IP.
This antibody is batch tested in ChIP.
We would recommend the customer to try IP only with formaldehyde fixation for 10min as we do in-house.
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.
Mr. Dan Stummer
Verified customer
投稿 Mar 15 2011