Flow Cytometry Analysis Protocol. Springer Lab - Harvard. Background on flow cytometry technique: Flow cytometry is a method which uses instrumentation scanning single cells flowing past excitation sources in a liquid medium. - [Read Flow Cytometry Analysis Protocol]
Protocol describes the growth and concentration of the alga Chlorogonium elongatum as a food source for culturing freshwater hypotrichs. Most freshwater hypotrichs (including Oxytricha nova, O. fallax, and O. trifallax; Euplotes aediculatus and E. eurystomous; and Stylonychia lemnae) can be grown to high density with Chlorogonium as the food organism. A similar regimen can be used to prepare other food sources such as Tetrahymena or bacteria (e.g., Aerobacter aerogenes). - [Read Growth and Concentration of Chlorogonium for Culturing Freshwater Hypotrichs Protocol]
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]
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]
Article presents an introduction to fluorescence microscopy. Includes: Fundamentals of Excitation and Emission; Stokes' Shift; Fading, Quenching, and Photobleaching; Fluorescence Light Sources; Filter Terminology; The Fluorescence Light Budget; Detecting Single Molecules. - [Read Introduction to Fluorescence Microscopy]
Isolation of poly A+ RNA from Total RNA by Oligo(deoxythymidine)cellulose Chromatography. Total RNA is first isolated from the tissues or cells and then mRNA is isolated by PolyA+ selection using oligo(dT) cellulose. This is necessary for all tissue sources rich in RNase (and cell lines). Lazo Lab - [Read Isolation of poly A+ RNA from Total RNA by Oligo(deoxythymidine)cellulose Chromatography]
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]
This protocol describes the storage of "homemade" purified antibodies. Antibodies from commercial sources are normally supplied with proper storage conditions. - [Read Storage of Purified Antibodies]
Under-Agarose assay is a useful method for observing the response of a cell population to one or more chemoattractant sources. BioProtocol - [Read The Under-Agarose Migration Assay]
DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.