Application
ChIP
Sample
Human Cell lysate - nuclear (Murine Neuroblastoma cells (Neuro 2A))
Negative control
Mock- Neuro 2A cells transfected with empty vector (pGL4.24 [luc/min] were processed in the same manner and considred as the negative control
Specification
Murine Neuroblastoma cells (Neuro 2A)
Detection step
Semiquantitative PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 16% w/v Paraformaldehyde
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 16% w/v Paraformaldehyde
Positive control
Input- ICP0 D enhancer fragment tansfected with human KLF15 expression vector. Cells were cross linked wth paraformaldehyde and lysed in FA lysis buffere. Samples were sonicated to produce 200-700bp fragments and precleared with salmon sperm slurry. Finally the samples were decrosslinked and the endpoint PCR was performed.
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: RIPA buffer
Dilution
1/100
Additional data
Additional Notes
FA lysis buffer-50 mM HEPES KOH, pH 7.5, 140 mM NaCl, 1 mM EDTA [pH 8.0], 1% Triton X-100, 0.1% sodium deoxycholate, 0.1% SDS
Protease inhibitors-Thermo Scientific Halt TM protease inhibitor cocktail 100X single use
The other antibodies used in the same samples are-Isotype (Is)ab,Glucocorticoid receptor (GR) ab (Diagram legend)
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提交于 Dec 14 2021