Simple ChIP Enzymatic Chromatin IP Kit (Millipore, USA) was used for ChIP assays according to the manufacturer's protocol. Cells were cross-linked with formaldehyde and collected in lysis buffer. Cell lysates were then sonicated to generate chromatin fragments of 200–300 bp and immunoprecipitated with EZH2 and H3K27me3-specific antibodies (CST) or IgG as control. Immunoprecipitated DNA was amplified by PCR using primers, which are listed in Supplementary Table 1 . EZH2 antibodies were obtained from Abcam. The H3 trimethyl Lys 27 antibody was purchased from Millipore. Precipitated chromatin DNA was recovered and analyzed by qRT-PCR.
Simple chip enzymatic chromatin ip kit
Manufactured by Merck Group
Sourced in United States
The Simple ChIP Enzymatic Chromatin IP Kit is a laboratory tool designed for the isolation and purification of chromatin-bound DNA fragments. It utilizes enzymatic digestion to fragment the chromatin, enabling the subsequent immunoprecipitation and recovery of DNA regions associated with specific proteins or histone modifications.
Automatically generated - may contain errors
3 protocols using simple chip enzymatic chromatin ip kit
1
ChIP Assay for Epigenetic Regulation
2
Chromatin Immunoprecipitation of TFEB
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Podocytes treated with BSA or AGE–BSA were used for ChIP assays. The Simple ChIP Enzymatic Chromatin IP Kit (Millipore, Billerica, MA, USA) was used following the manufacturer's protocol. Anti‐TFEB antibody (supplementary material, Table S3) was used to pull down DNA–protein complexes, and goat IgG was used as a control. Purified DNA was quantified using quantitative PCR (qPCR); primer sequences (TaKaRa Biotechnology, Dalian, PR China) are listed in the supplementary material, Table S4.
3
Chromatin Immunoprecipitation of Chondrocyte Transcripts
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PHCs were grown to confluency in 100-mm dishes, treated for 24 h with 50 nM miR-193b-3p or miR-Control, and used for ChIP assay analysis. The Simple ChIP Enzymatic Chromatin IP Kit (Millipore) was used following the manufacturer's recommended protocol. An anti-ac-H3 antibody (Millipore) was used to pull down DNA-protein complexes, and mouse IgG was used as a negative control. Purified DNA was quantified by qPCR. The sequences of primers used to amplify the SOX9, COL2A1, AGGRECAN, and COMP promoters are provided in Supplementary Material . The percent input method was used to identify enrichment.
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