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Past Issue in 2012
Volume: 2, Issue: 11
Cancer Biology
Propidium Iodide Staining of Cells for FACS Analysis
Fluorescence Activated Cell Sorting (FACS) is used to study DNA cell content. Propidium iodide (PI) intercalates into double-stranded nucleic acids and fluoresces. It is excluded by viable cells but can penetrate cell membranes of dying or dead cells. Thus PI staining is included in immunofluorescent staining protocols to identify dead cells. DNA staining can be used to study the cell cycle. Relative DNA content shows the proportion of cells in G1, G2 and S phases. Apoptotic cells show characteristic smear on DNA staining. Here a protocol to stain cells by PI is described.
Immunology
Isolation of Circulating Immune Complexes from TB Patient Serum for Serodiagnosis
Estimation of circulating immune complex in tuberculosis patients has shown better insight to the infection. Isolating circulating immune complex helps quantifying both antigen and antibody in the serum. It’s a simple procedure to improve the sensitivity and specificity in serodiagnosis of tuberculosis. This protocol may be modified to detect antigen/antibody in other infectious diseases.
Microbiology
In vitro Protein Kinase Assay Using Yeast Sch9
This protocol will describe experimental procedures for an in vitro kinase assay of the yeast protein kinase Sch9. This protocol can be tailored to detect kinase activity of other yeast protein kinase.
Neuroscience
Immunofluorescence Staining on Mouse Embryonic Brain Sections
This protocol comprises the entire process of immunofluorescence staining on mouse embryonic brains, starting from tissue preparation to mounting of the tissue sections.
Socially Transmitted Food Preference (STFP) Task Protocol
Temporally-graded retrograde amnesia (TGRA) refers to a phenomenon of premorbid memory loss whereby information acquired recently is more impaired than information acquired more remotely. Studies of human amnesia have illuminated this phenomenon (Hodges, 1994; Squire and Alvarez, 1995), but such studies necessarily rely on retrospective methods and imperfect tests. Studies in experimental animals have the advantage that retrograde amnesia can be studied prospectively, the locus and extent of brain lesions can be determined accurately, and the timing and strength of original learning can be precisely controlled. The socially transmitted food preference (STFP) task has been one of the most productive rodent behavioral tasks to study TGRA (Clark et al., 2002).
Stem Cell
Synchronize Human Embryonic Stem Cells at Different Cell Cycle Stage
Pluripotency and the capability for self-renewal are essential characteristics of human embryonic stem cells (hESCs), which hold great potential as a cellular source for tissue replacement. Short cell cycle (15-16 h) compared to somatic cells is another property of hESCs. Efficient synchronization of hESCs at different cell cycle stages is important to elucidate the mechanistic link between cell cycle regulation and cell fate decision. This protocol describes how to establish synchronization of hESCs at different cell cycle stages.