WebAug 18, 2024 · The promoter pull-down method takes an unbiased, biochemical approach to identify proteins bound to a defined promoter. Proteins are captured by flowing cellular lysate over promoter-bound beads. ... The streptavidin-biotin interaction is unstable at 70 °C, and thus this incubation allows complete elution of the DNA probe and all remaining ... WebJul 4, 2024 · National Center for Biotechnology Information
Biotinylated peptide: how to perform pull down?
WebNov 20, 2024 · Pulldown assay is a conventional method to determine protein-protein interactions in vitro. Expressing a protein of interest with two different tags allows testing whether both versions can be captured via one of the two tags as homooligomeric complex. This protocol is based on streptavidin b … WebFor biotin-based in vitro pull-down assay, GFP, PYL5 and eight polyketide cyclase-like proteins fused at the C-terminus to the HPB tag were purified from N. benthamiana as described above and stringently washed to remove bound endogenous metabolites [four washes with extraction buffer (150 m m NaCl, 50 m m TRIS-HCl pH 7.5, 10% glycerol, … f irving and sons
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WebCircRNA pull-down assay. A biotin-labeled circPTPRA probe was designed and synthesized by RiboBio Co., Ltd. The RNA pull-down assay was performed according to the instruction of the Pierce™ Magnetic RNA–Protein Pull-Down Kit (Thermo Scientific). The probe sequences used for the RNA pull-down assay are summarized in Additional … WebPull-down assays are useful for confirming the existence of protein-protein interaction. The interaction can be identified by other research techniques such as co-immunoprecipitation. The pull-down assay can be … WebRNA pull-down assay using nuclear extract 1. Biotin-labeled RNAs were in vitro transcribed with the Biotin RNA Labeling Mix (Roche) and T7 RNA polymerase (Promega), treated with RNase-free DNase I (Promega) and purified with RNeasy Mini Kit (QIAGEN). 2. Biotin-HRP Northern blot was followed by manufacture’s manual (NorthernMax kit, fir verification online