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Extraction and Solubilization of Total Protein from Microorganisms Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4224

Category: Bacterial Cell Culture Protocols

The growth conditions of microbial cell cultures and the time of sample collection should be optimized and standardized when growing cells for protein extraction. Because cells may excrete proteases and other extracellular enzymes, and compounds in the medium may interfere with extraction, wash cultures with an isotonic buffer, such as PBS or sucrose before solubilization. - [Read Extraction and Solubilization of Total Protein from Microorganisms Protocol]

Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of Intact DNA from Yeast - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3235

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol for preparation of DNA for pulsed-field gel electrophoresis: isolation of intact DNA from yeast. Yeast cells are first treated enzymatically to break down the cell walls and then resuspended in low-melting-temperature agarose plugs. The DNA is liberated by infusing the plugs with lysis buffer and proteases. This method is used to prepare both conventional and artificial yeast chromosomes. - [Read Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of Intact DNA from Yeast]

Recombinant aequorin and green fluorescent protein as valuable tools in the study of cell signalling - http://www.biochemj.org/bj/355/0001/3550001.pdf

Category: Signalling Signal Transduction Protocols

CHIESA et al 2001. J Biochem. Calcium-sensitive photoprotein aequorin used to analyse Ca2+ homoeostasis at the subcellular level. GFP chimaeras for studying the recruitment of protein kinase C isoforms and the activity of intracellular proteases. - [Read Recombinant aequorin and green fluorescent protein as valuable tools in the study of cell signalling]

Unmasking Hidden Epitopes with Proteases Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4528

Category: Immunohistochemistry Protocols: Antigen Retrieval Protocols

Fixation can mask epitopes. However, it is often possible to re-expose them using a gentle incubation with proteases, which removes obstructing structures and allows antibody access, as described here. Many proteases can be used for this procedure, including very crude preparations of proteases, such as pronase. However, using a better-characterized protease, such as trypsin, allows a more controlled reaction and better comparison between experiments. - [Read Unmasking Hidden Epitopes with Proteases Protocol]