107 cells were crosslinked with 1% formaldehyde at room temperature for 10 min. Chromatin was extracted and then sonicated to an average size of 300–1,000 bp. Immunoprecipitation was carried out by using Magna ChIP™ kit as recommended by the manufacturer (Millipore) and then purified using QIAquick PCR purification kit (Qiagen). Antibodies used (about 5 μg per 10–30 μg of DNA) were: Anti YY1 (H-414, Santa Cruz), Anti Trim28 (Anti tif1b- MAB3662, Millipore), Anti-trimethyl-Histone H3 (Lys9) (07–442, Millipore), Anti-trimethyl-Histone H3 (Lys27) (07–449, Millipore) and Anti-trimethyl-Histone H3 (Lys4) (07–473, Millipore). IP with IgG antibody (sc-2027, Santa Cruz) resulted in enrichment level < 1. Amplification was carried out by real-time PCR, and the bound/input values were then normalized by setting the negative control gene results to 1. Multiple assays of the same sample or the same gene sequence were analyzed in separate immunoprecipitations. All immunoprecipitations were repeated at least 3 times. Primer sequences used for qPCR are listed in Additional file 4 : Table S1. In Figure 6 D,E bound/input ratios were normalized to the U5-PBS primers value of the same sample and results are presented relative to the value in the negative sorted cells fraction.
Anti yy1 h414
Manufactured by Santa Cruz Biotechnology
Anti-YY1 (H414) is a laboratory reagent produced by Santa Cruz Biotechnology. It is a primary antibody that recognizes the YY1 (Ying Yang 1) protein. YY1 is a transcription factor involved in various cellular processes. This antibody can be used to detect and study the YY1 protein in samples through techniques such as Western blotting, immunoprecipitation, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Lightcycler 96, by Roche (2 mentions)
Anti trimethyl histone h3 lys9, by Merck Group (1 mentions)
Igg antibody sc 2027, by Santa Cruz Biotechnology (1 mentions)
Anti trimethyl histone h3 lys27, by Merck Group (1 mentions)
Anti trimethyl histone h3 lys4, by Merck Group (1 mentions)
Magna chip kit, by Merck Group (1 mentions)
Qiaquick pcr purification kit, by Qiagen (1 mentions)
3 protocols using anti yy1 h414
1
ChIP-qPCR Analysis of Chromatin
2
ChIP-seq and ChIP-qPCR for YY1 Binding
3
ChIP-seq and ChIP-qPCR for YY1 Binding
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The ChIP-seq data were compiled from the UCSC website (http://genome.ucsc.edu/ ). ChIP-qPCR was carried out according to the Upstate Biotechnology protocol. Approximately 10 × 106 HPC7 cells were used for each immunoprecipitation. Anti-YY1 (H414, Santa Cruz Biotechnology) antibody was used followed by protein A conjugated bead precipitation. Pre-immune controls used normal rabbit IgG. DNA samples were analyzed by real-time PCR using a Roche Lightcycler 96. Relative enrichments for each region were presented as the percentage of input. The primers for specific DNA sequences are listed in Table S1 .
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