Mammalia

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    Protocols in Current Issue
    Assaying the Effects of Splice Site Variants by Exon Trapping in a Mammalian Cell Line
    Authors:  Stuart W. Tompson and Terri L. Young, date: 05/20/2017, view: 5863, Q&A: 1
    [Abstract] There are several in silico programs that endeavor to predict the functional impact of an individual’s sequence variation at splice donor/acceptor sites, but experimental confirmation is problematic without a source of RNA from the individual that carries the variant. With the aid of an exon trapping vector, such as pSPL3, an investigator ...
    Measuring Procaspase-8 and -10 Processing upon Apoptosis Induction
    Authors:  Sabine Pietkiewicz, Clara Wolfe, Jörn H. Buchbinder and Inna N. Lavrik, date: 01/05/2017, view: 5331, Q&A: 0
    [Abstract] Apoptosis or programmed cell death is important for multicellular organisms to keep cell homeostasis and for the clearance of mutated or infected cells. Apoptosis can be induced by intrinsic or extrinsic stimuli. The first event in extrinsic apoptosis is the formation of the Death-Inducing Signalling Complex (DISC), where the initiator caspases-8 ...
    Quantification of T Cell Antigen-specific Memory Responses in Rhesus Macaques, Using Cytokine Flow Cytometry (CFC, also Known as ICS and ICCS): Analysis of Flow Data
    Authors:  Andrew W. Sylwester, Scott G. Hansen and Louis J. Picker, date: 04/20/2014, view: 11184, Q&A: 0
    [Abstract] What was initially termed ‘CFC’ (Cytokine Flow Cytometry) is now more commonly known as ‘ICS’ (Intra Cellular Staining), or less commonly as ‘ICCS’ (Intra Cellular Cytokine Staining). The key innovations were use of an effective permeant (allowing intracellular staining), and a reagent to disrupt secretion (trapping cytokines, thereby enabling ...
    Quantification of T Cell Antigen-specific Memory Responses in Rhesus Macaques, Using Cytokine Flow Cytometry (CFC, also Known as ICS and ICCS): from Assay Set-up to Data Acquisition
    Authors:  Andrew W. Sylwester, Scott G. Hansen and Louis J. Picker, date: 04/20/2014, view: 10943, Q&A: 0
    [Abstract] What was initially termed ‘CFC’ (Cytokine Flow Cytometry’) is now more commonly known as ‘ICS’ (Intra Cellular Staining), or less commonly as ‘ICCS’ (Intra Cellular Cytokine Staining). The key innovations were use of an effective permeant (allowing intracellular staining), and a reagent to disrupt secretion (trapping cytokines, thereby enabling ...
    Cell Surface Protein-protein Binding on COS-7 Cells
    Authors:  Kae-Jiun Chang and Matthew N. Rasband, date: 01/05/2014, view: 7060, Q&A: 0
    [Abstract] Examination of interactions between a transmembrane protein and a soluble protein by pull-down or immunoprecipitation assays can be tricky and complicated due to the detergent extraction of membrane proteins during the lysate preparation step. The choice and concentration of detergents must be determined empirically and the procedure can be ...



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