Transient transfection into mammalian cells is a convenient way to over express and obtain protein expression. Protocol includes: Culture conditions; Transfection of experimental cells; Preparation of Mammalian Cell Lysate for Luciferase assay. - [Read Transient Transfection and Luciferase Assay Protocol]
In this stage of the protocol, a mammalian cDNA library constructed in a plasmid such as pJG4-5 is transformed into yeast strains containing pBAIT and the lexAop-lacZ reporter plasmid. PJG4-5 expresses the cloned cDNAs from a cassette containing a transcriptional activation domain and other moieties under the control of the yeast GAL1 promoter. - [Read Two-hybrid Systems Stage 2: Selecting an Interactor Protocol]
Ultimate His-UB Assay for Mammalian Cells. Tansey Lab Protocols. William P. Tansey. PREPARATION OF NI-NTA-AGAROSE, HARVESTING THE TRANSFECTION., ELUTION OF HIS-TAGGED PROTEINS. - [Read Ultimate His-UB Assay for Mammalian Cells]
A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.