Application
ChIP
Sample
Mouse Tissue lysate - nuclear (Thymus)
Negative control
upstream element of Gata6
all the procedures were done in wild type and in Satb1 knockout cells
Specification
Thymus
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Positive control
RHS6 region (previously worked in our laboratory)
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: ChIP Dilution Buffer
Additional data
Additional Notes
ABTRIAL TESTING:
20 million of freshly prepared mouse thymocytes from each C57BL/6 wild type and Satb1 fl/fl CD4Cre+ conditional knockout was fixed with formaldehyde for 10 min. Cells were lysed and purified using 5-8 M urea gradient. Chromatin was sonicated to get a bulk of sheared chromatin bellow 500 bp. The ChIP was performed with 20 µl of Dynabeads® Protein G pre-incubated with antibodies. Two titrations of 1.2 µg (left in the figure) and 2.5 µg (right in the figure) Abcam ab109122 antibodies (EPR3951, Lot: GR137720-3) were used for both wt and knockout. The immunoprecipitated DNA was analyzed for RHS6 region as a positive control (PC based on previous results of our laboratory) and upstream element of Gata6 as a negative control (NC).
Abcam response
Thank you for providing results of your testing. I am sorry that the antibody - not yet tested or guaranteed in ChIP - has failed. Nevertheless, if of interest, we encourage further Abtrial tests.
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
Submitted Jul 11 2016