ChIP assay was conducted by use of the EZ ChIP™ Chromatin Immunoprecipitation Kit for cell line (Millipore, Bedford, MA, USA). VCaP and LNCaP cell lines were first fixed in 1% formaldehyde for 15 min crosslink. Thereafter, crosslinked chromatin was ultra-sonicated into 200- to 1000-bp fragments, and then immunopreciated with anti-PAX5 or control anti-IgG bound-protein G beads for 6 h. qRT-PCR was used to evaluate the enrichments of DNA fragments immunoprecipitated by antibodies. ChIP assay was performed in triplicate.
Ez chip chromatin immunoprecipitation kit for cell line
Manufactured by Merck Group
Sourced in United States, United Kingdom
The EZ ChIP™ Chromatin Immunoprecipitation Kit is a laboratory equipment product designed for chromatin immunoprecipitation (ChIP) analysis of cell lines. It provides the necessary reagents and protocols for isolating and purifying DNA-protein complexes from cell samples.
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3 protocols using ez chip chromatin immunoprecipitation kit for cell line
1
ChIP Assay for PAX5 Binding
2
ChIP Assay for Transcription Factor Binding
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ChIP assay was implemented by use of the EZ ChIP™ Chromatin Immunoprecipitation Kit for cell line (Millipore, Bedford, MA). U87 or U251 cell samples were initially fixed in the 1% PFA for 15 min’s crosslink then processed by ultrasonic for cutting DNA into fragments (500 bp). Following that, the 6 h of immunoprecipitation was undertaken with antibodies against control IgG and YY1, respectively. 30 μl of magnetic beads were added afterwards to collect the precipitated chromatin for RT-qPCR analysis.
3
ChIP Assay for Cell Line
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ChIP assay was performed in light of the protocol of EZ ChIP™ Chromatin Immunoprecipitation Kit for cell line (Millipore). The treated cells were fixed in formaldehyde for crosslink and sonicated for shearing DNA, followed by incubation with indicated antibodies for 6 h. Normal mouse IgG acted as the NC. The precipitates from beads were analyzed by RT-qPCR.
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