lysates Protocols

Category: Chromatography Protocols: Affinity Chromatography Protocols

Recombinant protein or a chemically synthesized bioactive fragment is immobilized on resin and used as a probe to capture interacting proteins directly from a cell extract.  Affinity-purified proteins are fractionated by gel electrophoresis and visualized by Coomassie staining.  Proteins that interact specifically are identified by comparing this gel profile to one obtained from cell lysates passed over a control resin lacking the immobilized probe protein. - [Read Affinity Purification of Interacting Proteins from Cell Lysates Protocol]

Category: Signalling Signal Transduction Protocols

Protocol allows you to measure the content of cyclic adenosine 3',5'-monophosphate (cyclic AMP or cAMP) in splenic B lymphocytes (B cells) in an enzyme-linked immunoassay. This protocol utilizes acetylation of cAMP to improve sensitivity and reduce interference. Protocol includes information on: how to determine cAMP, calculations and reagents and materials. - [Read Assay of Cyclic AMP in Lysates of Cells]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol describes the DNA content of bacteriophage lambda stocks can be easily and rapidly estimated. - [Read Assaying the DNA Content of Bacteriophage lambda Stocks and Lysates by Gel Electrophoresis Protocol]

Category: Protein Protocols: Western Blot Protocol

How to prepare cell lysates for western blotting. - [Read Cell Lysates For Western Blotting]

Category: Molecular Biology Protocols: Phage Protocols and Methods: Lambda Phage Protocols

Column Method for Mini-prep of lambda phage DNA purification from lysates. Donis Keller Lab - [Read Column Method for Lambda Phage DNA Preparation]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

Mini-prep method for lambda phage DNA purification from lysates. - [Read Column Method for Lambda Phage DNA Preparation Protocol]

Category: Xenopus Protocols: Xenopus Embryo Manipulation Protocols

Protocol for embryo lysates and immunoprecipitation: protein A agarose & protein A magnetic beads. Includes: Embryo lysates; Immunoprecipitation with protein A agarose; Immunoprecipitation with protein A magnetic beads for silver stain/sequencing analysis. - [Read Embryo Lysates & Immunoprecipitation Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Freezing and Thawing Eukaryotic Cells in liquid nitrogen, Freezing Cell Lysates for Western Blots, Thawing Cells from Liquid Nitrogen. Dr. Bart Frank Oklahoma. - [Read Freezing and Thawing Eukaryotic Cells]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

A crude lysate gel assay can be performed to roughly quantitate the DNA in lysates. This is often a valuable time saving step to determine if the phage yield is sufficient to warrant continuing the procedure. - [Read Gel Assay to Determine DNA Content of Phage Lysates Protocol]

Category: Western Blot Protocols: General Western Blot Methods Protocols

For immunoblotting experiments, it is often important to compare the total amount of an antigen from many different sources or to learn if a particular source has the antigen under study.  In the approach described here, tissue cultures, bacteria, yeast cells, tissues, and other sources of antigens are disrupted directly in an electrophoresis sample . - [Read Immunoblotting: Preparing Cell Lysates Protocol]

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol for immunoprecipitation of endogenous MEKK1 and kinase assay. Includes: Preparation of lysates; Immunoprecipitation; Kinase assay. - [Read Immunoprecipitation of Endogenous MEKK1 and Kinase Assay Protocol]

Category: Immunology: Immunoprecipitation Protocols

For many sources of antigens, one useful method of lysis is to treat cells with harsh, denaturing solutions to release most of the protein antigens, as described here. The lysates are then diluted to reduce the denaturing conditions to levels that are suitable for the formation of antibody-antigen complexes. The resulting solution is precleared prior to immunoprecipitation. - [Read Immunoprecipitation: Denaturing Lysis Protocol]

Category: Protein Protocols: Protein Trafficking

Import assays into chloroplasts. Includes: Preparation of chloroplasts and lysates; Transport of lumenal proteins. - [Read Import Assays Into Chloroplasts: Integration or Transport into Thylakoids]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

5 ml liquid lysates are prepared when a small amount of DNA from a large number of lambda clones is needed. The lysates can be made using 10- 20 ul of a stock lysate or a 100-fold amplified phage "macroplaque" as the inoculum. - [Read Liquid Phage Lysates Protocol]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

To prepare phage lysates to be used for small or large scale phage DNA preps. This method usually produces lysates with titers of 2-8x10e10 pfu/ml. - [Read Phage Plate Stock Lysates Protocol]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

Protocol for the isolation of single bacteriophage lambda plaques to be used for making lysates. - [Read Picking Single Phage Plaques Protocol]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

Bacteriophage {lambda} particles are recovered from bacterial lysates by precipitation with polyethylene glycol at high ionic strength. - [Read Precipitation of Bacteriophage {lambda} Particles from Large-scale Lysates Protocol]

Category: Signalling Signal Transduction Protocols

This protocol provides a sufficient sample for several determinations of cAMP using the acetylation protocol. The method chosen for measuring the content of cyclic adenosine 3',5'-monophosphate (cyclic AMP or cAMP) in splenic B lymphocytes (B cells) is an enzyme linked immunoassay system. Protocol includes information on: Treatment of Cells and Preparation of Extracts; Reagents and Materials. - [Read Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination]

Category: Signalling Signal Transduction Protocols

Protocol provides a method of achieving a sufficient sample for several determinations of cAMP using Amersham’s acetylation protocol. Protocol includes information on: Treatment of Cells and Preparation of Extracts; Reagents and Materials. - [Read Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination for Dual Ligand Screen]

Category: Immunology: B Cell Protocols

The method chosen for measuring the content of cyclic adenosine 3',5'- monophosphate (cyclic AMP or cAMP) in splenic B lymphocytes (B cells) is an enzymelinked immunoassay system. Includes: Treatment of Cells and Preparation of Extracts. - [Read Preparation of B-Lymphocyte Lysates for Cyclic AMP Determination for Dual Ligand Screen Protocol]

Category: Protein Protocols: Protein Extraction From Cells

Preparation of cell lysates from yeast by enzymatic digestion of the cell wall. - [Read Preparation of cell lysates from yeast by enzymatic digestion of the cell wall]

Category: Protein Protocols: Protein Extraction From Cells

Preparation of cell lysates from yeast using a French Press. he Protein Expression and Purification Facility, EMBL - [Read Preparation of cell lysates from yeast using a French Press]

Category: Protein Protocols: Protein Extraction From Cells

Preparation of cell lysates from yeast using glass beads vortexing. Includes lysis buffer recipes. The Protein Expression and Purification Facility, EMBL - [Read Preparation of cell lysates from yeast using glass beads vortexing]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

In this protocol, a bacterial lysogen is constructed from a recombinant bacteriophage {lambda} encoding a fusion protein of interest. The resulting lysogenic colonies are induced to synthesize the fusion protein, which is then isolated in preparation for functional and biochemical analyses. - [Read Preparation of Lysates Containing Fusion Proteins Encoded by Bacteriophage {lambda} Lysogens]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

This rapid method is used to screen bacteriophage {lambda}gt11 clones for the production of immunodetectable fusion proteins. After optimizng the conditions of infection and induction, the method can be used to produce preparative amounts of a fusion protein. - [Read Preparation of Lysates Containing Fusion Proteins Encoded by Bacteriophage {lambda}: Lytic Infection]