Application
ChIP
Sample
Human Cell lysate - nuclear (HUVEC)
Negative control
olfactory region 8 (OR8)
Specification
HUVEC
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 60 minute(s) and 0 second(s)
Specification of the cross-linking agent: 20 min EGS + 40 min with PFA w
Duration of cross-linking step: 60 minute(s) and 0 second(s)
Specification of the cross-linking agent: 20 min EGS + 40 min with PFA w
Positive control
5’ prime UTR of ZNF335 and HIST2H2AC genes
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: Shearing buffer
Additional data
Additional Notes
Chip assay was performed using 25million HUVECs and Chip It High Sensitivity kit (Active Motif) and Abcam HMGB2 antibody (ab 124670). The cells were crosslinked with 15mM ethylene glycol bis(succinimidylsuccinate) (EGS) for 20 minutes at room temperature, followed by the addition of Complete Cell Fixative Solution for 40 minutes at 4oC provided by the kit. 25µl of Abcam HMGB2 antibody were added in 25µg of sheared chromatin. QPCR was performed using positive primers against the 5’ prime UTR OF ZNF335 and HIST2H2AC genes and negative primer against the olfactory region 8. Figure 1 shows the relative amplification of positive controls against the negative.
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. Theodore Georgomanolis
Verified customer
提交于 May 25 2016