Project description:In vitro pull down assay was performed using recombinant GST and GST fusion SOX2 protein to precipitate total RNA from urothelial carcinoma cell line BFTC905 Unbound RNA from both reactions and matrix-associated RNA from SOX2 pull down reaction was extracted using TRIZOL reagent
Project description:In vitro pull down assay was performed using recombinant GST and GST fusion SOX2 protein to precipitate total RNA from urothelial carcinoma cell line BFTC905 Unbound RNA from both reactions and matrix-associated RNA from SOX2 pull down reaction was extracted using TRIZOL reagent This experiment is to identify mRNA bound by SOX2 under in vitro experimental condition
Project description:The TET3 CXXC domain has unique DNA binding properties. It binds to DNA in a cytosine-dependent manner that prefers binding to CpG dinucleotides but is not restricted by the CpG-content, distinct from other well-characterized CXXC domains. To map the TET3 CXXC domain binding sites across the human genome, we purified the GST-tagged TET3 CXXC domain protein and performed the GST pull-down assay using the genomic DNA purified from HEK293T cells. The enriched DNA fragments were then sequenced and aligned to human genome(hg19). We used the GST pull-down assay followed by DNA deep sequencing to map the DNA bound by the TET3 CXXC domain in vitro.
Project description:A pull-down assay by incubating the His-tagged PPARalpha-LBD with various brain tissue (cortex, cerebellum, and hippocampus) extracts.
Project description:Genome-wide identification of R-loops almost uniquely relies on antibody based pull-down, a method that shows sequence and structural biases. Here we describe a novel approach – DREAM-seq – centered around the use of endonuclease digestions. Unstimulated and stimulated B cells showed that R-loops are more widespread than anticipated, mostly covering all actively transcribed genes.
Project description:HOIP truncations proteins GST-PUB, GST-NZF, GST-UBA and GST-RBR-LDD were purified, GST was used as a negative control. The tissue lysates from the C57BL/6 mice were incubated with glutathione-S-transferase (GST) beads which bound to the GST-HOIP truncation proteins beforehand. The pull-down component were separated by SDS-PAGE and stained with Coomassie brilliant blue. The band were cut off and subject to mass spectrometry.