Protein-protein interaction assay with GST fusion proteins
GST fusion proteins, as recombinant proteins expressed in bacteria, have been used in thousands of research projects since their introduction (Smith and johnson
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Amplification and storage of mucoid libraries (amplification on filter membranes)
Applying this amplification method, the library is not prone to distortion, due to the fact that mixed colonies containing different recombinant mucilages do no
Experiments with two-hybrid and other two-component systems
This experiment describes experimental methods regarding two-hybrid and other two-component systems. This experiment comes from the next volume of the Laborator
Amplification and storage of mucoid libraries (amplification in liquid medium)
Do not over-amplify the mucoid library, as this will inevitably cause distortion of the original genome. Faster-growing clones will be over-presented, unstable
Ecdysone regulation induces cellular gene expression experiments
The following protocol is modified from the protocol that accompanies the Ecdysone-Induced Mammalian Expression System (Invitrogen); a similar system is sold by
GFP and fluorescence resonance energy transfer techniques for protein interaction assays
We have divided the protocol into three phases: the first phase describes the preparation of proteins and the labeling of proteins with fluorescent dyes; the se
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
Analysis of interacting proteins using BIAcore
BIAcore surface plasmon resonance is widely used for (1) studying the interaction process between various biomolecules (e.g., peptides, proteins, oligonucleotid
Isolation of BAC DNA from small cultures
Small amounts of BAC DNA were prepared from 5 ml of BAC-transformed cell cultures.The DNA was prepared by alkaline lysis.The yield of BAC DNA can be as high as