Application
ChIP
Sample
Human Cell lysate - whole cell (MCF-7 nuclear and cytosolic lysates)
Negative control
A nearby region of the genome that is not a known enhancer in MCF-cells
Specification
MCF-7 nuclear and cytosolic lysates
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: 1% formaldehyde at room temper
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde at room temper
Other product details
Incubation time
4 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: ChIP Dilution buffer
Additional data
Additional Notes
Histone H3 antibody was used as a control to measure the relative fold occupancy for the H3K4me1 and H3K27ac on enhancer and a nearby control region.
ChIP Dilution buffer: 50mM Tris (pH7.4), 10 mM HEPES-KOH (pH7.5), 100 mM NaCl, 1 mM EDTA pH8, 0.5% Triton X-100, 0.1% Sodium Deoxycholate, 0.1% SDS.
2 microliter of antibody used per ChIP
Overnight incubation with Protein A/G beads, 4h after addition of antibody.
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提交于 May 08 2018