location Protocols

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

Protocol describes systems in which the membranes are allowed to float through a density gradient from a dense load zone. This is an ideal format. - [Read A Self-generated Gradient to Discriminate a Cytosolic or Membrane Location of Large Protein Complex]

Category: Immunology: Immunoprecipitation Protocols: Chromatin Immunoprecipitation Protocols

Genome-wide location analysis, also known as ChIP-Chip, combines chromatin immunoprecipitation and DNA microarray analysis to identify protein-DNA interactions that occur in living cells. Protein-DNA interactions are captured in vivo by chemical crosslinking. Cell lysis, DNA fragmentation and immunoaffinity purification of the desired protein will co-purify DNA fragments that are associated with that protein. - [Read Chromatin Immunoprecipitation and Microarray-Based Analysis of Protein Location Protocol]

Category: Stem Cell Protocols

This protocol describes a method for freezing and thawing ES cells using cryovials. It is important to freeze ES cell stocks as soon as possible to reduce the time that they are in culture. A careful record should be kept of the number of times cells are passaged and the location of the cryovials. - [Read Freezing and Thawing of Embryonic Stem (ES) Cells Using Cryovials Protocol]

Category: Molecular Imaging: Electron Microscopy Protocols

In brief, the reaction mixture, is prepared in exactly the same way as if the sample is to be stained or shadowed for electron microscopy, but once the... Gold labeling technique for electron microscopic identification and location of proteins. - [Read Gold labeling of proteins for electron microscopy]

Category: Plant Biology Protocols: Plant Genetics Protocols

To accurately predict the activity of a transgene it is critical to understand its location and dynamics in the 3-D interphase nucleus. Developed in situ methods to visualize transgenes (including single copy genes) & their transcripts during interphase from different tissues & plant species. These techniques reduce the time necessary for characterization of transgene integration by eliminating the need for time-consuming segregation analysis and extend characterization to the interphase nucleus - [Read In Situ Methods to Localize Transgenes and Transcripts in Interphase Nuclei]

Category: Microinjection Protocols: Microinjection of Proteins Protocols

Activation and inactivation of proteins using photoactivation of caged peptides or proteins offer insights into cellular dynamics not achievable using genetic means. The ability to selectively alter the activity of a specific protein at a defined time and location inside a cell allows the correlation of changes in protein activity and cellular behavior. A caged compound, peptide, or protein is prepared by covalently linking it to a photolabile, protecting group. - [Read Introduction of Caged Peptide/Protein into Cells Using Microinjection Protocol]

Category: Cytogenetics Protocols

Protocol describes a recently developed method — methylation-specific digital karyotyping (MSDK) — that enables comprehensive and unbiased genome-wide DNA methylation analysis. Using a combination of a methylation-sensitive mapping enzyme (for example, AscI) and a fragmenting enzyme (for example, NlaIII), short sequence tags can be obtained and uniquely mapped to genome location. - [Read Methylation-Specific Digital Karyotyping Protocol]

Category: Immunohistochemistry Protocols: Antibody Staining Protocols

Most histological studies are carried out on paraformaldehyde-fixed, paraffin-embedded tissue samples. Therefore, there is an extensive atlas of most tissues and organs prepared from these sources, and comparing the location of antigens to these data is immediately informative. The fixation and embedding procedures are harsh, however, and many antigens are not well preserved. - [Read Preparing Paraffin Tissue Sections for Immunostaining Protocol]

Category: Fungi Protocols: Aspergillus Protocols

Protocol allows the isolation and enumerate Aspergillus spp spores in air. Includes: Sampling Procedure; Sampling Location; Selection of Sampling Time; Sampling Steps; Laboratory Procedure; Enumerating the Colony Forming Units. - [Read Sampling of Aspergillus spp Spores in Air Protocol]

Category: DNA Protocols: SNP Protocols

SNP detection with mutagenic primers. Input sequence will be searched to find changes in one nucleotide near the location of the SNP, so that mutagenic primers may be easily designed. Bikandi, J., San Millán, R., Rementeria, A., and Garaizar, J. In silico analysis of complete bacterial genomes: PCR, AFLP-PCR, and endonuclease restriction. Bioinformatics 2004 Mar 22;20(5):798-9. - [Read SNP detection with mutagenic primers]