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Past Issues in 2012

Volume: 2 Issue: 23

Biochemistry

Glycophosphosphingolipid (GSPL) Purification Protocol Author: Tripti De, date: 12/05/2012, view: 7911, Q&A: 0

Glycosylated ceramide phosphorylinositol are present in many species of fungi and mushrooms and bacteria and parasitic organisms like leishmania. These are usually membrane raft associated and are not easily extracted by conventional methodologies. This extraction method gives higher yield of the glycolipid. Glycosphingolipids are usually ...

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Immunology

Viral Immunofluorescence with Rift Valley Fever Virus Infected MEFs in a 96 Well Plate Authors: Theresa S. Moser and Sara Cherry, date: 12/05/2012, view: 8970, Q&A: 0

Immunofluorescence is a method to detect viral infection in multiple types of host cells. This procedure can be adapted for both high-throughput and low-throughput assays for any virus for which there are antibodies available. Time of infection and virus multiplicity of infection (MOI) vary and should be optimized for each virus and host cell ...

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Adoptive Transfer of Isolated Bone Marrow Neutrophils Authors: Jimena Tosello Boari and Eva Acosta Rodríguez, date: 12/05/2012, view: 14715, Q&A: 0

Adoptive transfer experiments of specific cell populations are widely used methods to assess the role of the injected population on an ongoing process. In the last years, new and unprecedented roles in the regulation of immune responses have been reported for neutrophils. The following protocol is to be used to isolate neutrophils from bone marrow ...

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Standard 4-hours Chromium-51 (⁵¹Cr) Release Assay Author: Julie Gertner-Dardenne, date: 12/05/2012, view: 18763, Q&A: 0

⁵¹Cr-release assays are commonly used for the precise and accurate quantification of cytotoxicity, particularly in the study of tumor cytolysis. This test has the advantage of requiring only very few cells.

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Ex vivo Human Antigen-specific T Cell Proliferation and Degranulation Authors: Willemijn Hobo, Wieger Norde and Harry Dolstra, date: 12/05/2012, view: 11702, Q&A: 0

Proliferative capacity and degranulation are important features of antigen-specific CD8⁺ T cells. By combining tetramer staining with a CFSE staining, we were able to enumerate the total number of antigen-specific T cells, as well as their number of divisions upon antigen-specific stimulation during a week. In addition, we performed ...

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Neuroscience

A Protocol for Electrophoretic Mobility Shift Assay (EMSA) from Primary Neuron Author: Jiali Li, date: 12/05/2012, view: 23399, Q&A: 3

The interaction of transcriptional or co-transcriptional factors with DNA is crucial for changes of neuronal gene expression during normal brain development as well as neurodegeneration. The electrophoretic mobility shift assay (EMSA) is a very powerful technique for studying changes of neuronal gene expression and determining protein: DNA ...

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Plant Science

Plastic Embedding and Sectioning of Plant Tissues Author: Tie Liu, date: 12/05/2012, view: 13202, Q&A: 1

Plastic (resin) embedding provides exclusively improvements to cellular definition compared to paraffin embedding. The combination of strongly cross-linking paraformaldehyde with glutaraldehyde and post fixed with OsO₄ is the fixative of choice for high-resolution light microscopy and electron microscopy. For this reason, this method is ...

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Preparation of cDNA Library for dRNA-seq Authors: Feng Li and Barbara Baker, date: 12/05/2012, view: 12336, Q&A: 0

microRNAs (miRNAs) are ubiquitous regulators of gene expression in eukaryotic organisms, which guide Argonaute proteins (AGO) to cleave target mRNA or inhibit its translation based on sequence complementarity. In plants, miRNA directed cleavage occurs on the target mRNA at about 10 to 11 nucleotide (nt) up stream to the site where the 5’ end of ...

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Grafting of Potato Plants Author: Paula Suárez-López, date: 12/05/2012, view: 9034, Q&A: 1

Grafting is a very useful technique for testing the transmission of long-distance signals in plants and is used in agriculture and gardening for different purposes. This protocol, based on a previously published one (Jackson et al., 1998), describes a grafting method for potato plants, which has successfully been used to test the ...

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