Gel blocking assay of DNA binding proteins
The gel-blocking assay for analyzing DNA-binding proteins was invented by Fried and Crothers (1981) and was the first to provide a dynamic method for analyzing
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DNA transfection experiment using Polybrene
The Polybrene and DMSO transfection method described below is a modification of the method of Aubin et al. (1997). Using this method, plasmid DNA can be efficie
Experiments with tetracycline as a regulator of inducible gene expression
The following protocol is divided into 3 stages: stable transfection of fibroblasts with pTet-tTAk, stable transfection of NIH-3T3 cells inducible to express tT
Screening of unamplified mucoid libraries by hybridization (filter film photocopying)
Thick bacterial colonies transformed by mucilage can be screened by hybridization, a method derived from the large-scale screening of plasmid-transformed coloni
Determination of chloramphenicol acyltransferase content by thin-layer chromatography experiment
In this protocol, we list three different assays that can be used to determine CATase activity expressed after transfection of mammalian cells with the reporter
Determination of luciferase in mammalian cell extracts
The assay of luciferase in mammalian cell extracts can be used to (1) measure the amount of luciferase in animal tissues, and (2) measure other enzymatic activi
Conjugate transcription profiling experiments
Nuclear contig analysis is used to determine whether a gene is transcriptionally regulated in a given clone. Theoretically, nuclear contig analysis should be pe
GST fusion protein sedimentation assay
The GST sedimentation experiment has an advantage over far western (Scheme 2): the probe protein is incubated with a possible partner protein in a more natural
Immunoprecipitation assay to determine binding proteins
Many of the protein-protein interactions present in intact cells are maintained when cells are lysed under nondenaturing conditions. This rhopper can be used to