Molecular Biology

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    Immunophenotyping and Intracellular Staining of Fixed Whole Blood for Mass Cytometry (CyTOF)
    Authors:  Sangeeta Kowli and Holden Maecker, date: 09/05/2020, view: 543, Q&A: 0
    In this report, we present the implementation of mass cytometry for intracellular staining using fixed whole blood. In our assay described here, 250 µl of whole blood, is stimulated in vitro with PMA/ionomycin (or left unstimulated), in the ...
    Heterologous Expression and Purification of SARS-CoV2 Nucleocapsid Protein
    Authors:  Ankur Garg, Lihong Liu, David D. Ho and Leemor Joshua-Tor, date: 08/05/2020, view: 948, Q&A: 0
    This protocol describes a step by step method for heterologous expression of SARS-CoV2 Nucleocapsid (N) protein in Escherichia coli. Moreover, this protocol includes steps to purify the N protein to high purity and homogeneity. Thus, ...
    Heat Induced Epitope Retrieval (HIER) Assisted Protein Immunostaining in Maize
    Authors:  Katsutoshi Tsuda and George Chuck, date: 06/05/2019, view: 1309, Q&A: 0
    Protein immunostaining provides important spatio-temporal information about gene expression. Even using high quality antibodies, signal reproducibility and specificity can be problematic depending on tissue fixation methods. For example, ...
    Western Blot in Maize
    Author:  María Jazmín Abraham-Juárez, date: 06/05/2019, view: 3584, Q&A: 0
    Since its introduction in 1979 (Towbin et al., 1979), protein blotting (Western blot) has become a routine tool in molecular biology research. It is used to detect low amounts of proteins in complex samples or to monitor protein expression ...
    Protein Immunoprecipitation in Maize
    Author:  María Jazmín Abraham-Juárez, date: 06/05/2019, view: 1565, Q&A: 0
    Protein immunoprecipitation (IP) is a method used to identify cellular protein complexes. Whence, antibodies that specifically recognize a native bait protein or an epitope tag fused to the protein of interest allow the protein of interest to be ...
    High-throughput YO-PRO-1 Uptake Assay for P2X7 Receptors Expressed in HEK Cells
    Authors:  Akira Karasawa and Toshimitsu Kawate, date: 07/20/2018, view: 3988, Q&A: 0
    P2X7 receptors are extracellular ATP-gated ion channels that play broad physiological and pathological roles in animals (Sluyter, 2017). Activation of P2X7 receptors lead to the opening of membrane channels permeable for small cations like Na+ ...
    Overcoming Autofluorescence to Assess GFP Expression During Normal Physiology and Aging in Caenorhabditis elegans
    Authors:  Alina C. Teuscher and Collin Y. Ewald, date: 07/20/2018, view: 6544, Q&A: 0
    Green fluorescent protein (GFP) is widely used as a molecular tool to assess protein expression and localization. In C. elegans, the signal from weakly expressed GFP fusion proteins is masked by autofluorescence emitted from the intestinal ...
    In vitro Chaperone Activity Assay Using α-Amylase as Target Protein
    Authors:  Jeeshma Nambidi Parambath, Gayathri Valsala, Karthik Menon and Shiburaj Sugathan, date: 06/20/2018, view: 4188, Q&A: 0
    Small heat shock proteins (sHSP) are stress proteins which are ubiquitously found in almost all living organisms. They function as molecular chaperones, which assist in protein folding during translation and in the prevention of irreversible protein ...
    Activation of ER-regulated Fusion Proteins In Vivo
    Author:  Jason Reuter, date: 12/20/2011, view: 11208, Q&A: 0
    Regulatable protein fusions can be made by attaching the hormone binding domain of the estrogen receptor to the N or C-terminus of a protein of interest. Activation of such ER-fusion proteins in vivo can be achieved by daily administration ...
    Infiltration of Nicotiana benthamiana Protocol for Transient Expression via Agrobacterium
    Author:  Xiyan Li, date: 07/20/2011, view: 56076, Q&A: 3
    Transient expression in tobacco plant (Nicotiana benthamiana) is used to determine the subcellular location of a protein of interest when tagged with a reporter such as green fluorescent protein (GFP), or to mass produce proteins without ...



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