In vitro AMPylation/Adenylylation of Alpha-synuclein by HYPE/FICD
    作者:Ali Camara, Anwesha Sanyal, Sayan Dutta, Jean-Christophe Rochet and Seema Mattoo日期:09/20/2020,浏览量:83,Q&A: 0
    [Abstract] One of the major histopathological hallmarks of Parkinson’s disease are Lewy bodies (LBs) –cytoplasmic inclusions, enriched with fibrillar forms of the presynaptic protein alpha-synuclein (α-syn). Progressive deposition of α-syn into LBs is enabled by its propensity to fibrillize into insoluble aggregates. We recently described a marked reduction ...
    In vitro Assessment of Pathogen Effector Binding to Host Proteins by Surface Plasmon Resonance
    作者:Marina Franceschetti, Mark J. Banfield, Clare E. M. Stevenson and Juan Carlos De la Concepcion日期:07/05/2020,浏览量:1146,Q&A: 0
    [Abstract] The mechanisms of virulence and immunity are often governed by molecular interactions between pathogens and host proteins. The study of these interactions has major implications on understanding virulence activities, and how the host immune system recognizes the presence of pathogens to initiate an immune response. Frequently, the association ...
    Molecular Size Analysis of Recombinant Importin-histone Complexes Using Analytical Ultracentrifugation
    作者:Abhilash Padavannil, Chad A. Brautigam and Yuh Min Chook日期:05/20/2020,浏览量:941,Q&A: 0
    [Abstract] Histones constitute the protein components of nucleosomes. Despite their small sizes, histones do not diffuse through the nuclear pore complex. Instead, they are transported to the nucleus by importins, either alone or in complex with histone chaperones. Determining the molecular size of the importin-histone complexes is key to understanding the ...
    Fibrin Breakdown Assay
    作者:Jiayue Ling, Connor M. Blair and George S. Baillie日期:04/20/2020,浏览量:1395,Q&A: 0
    [Abstract] Fibrinolysis is an integral part of the matrix remodeling process that contributes to tissue repair. Fibrin clots are broken down during fibrinolysis in a controlled process. Fibrin degradation products (FDPs) have also been shown to have a role in the regulation of cell growth and are implicated in various vascular diseases. This protocol was ...
    Quantitative Determination of Ca2+-binding to Ca2+-sensor Proteins by Isothermal Titration Calorimetry
    作者:Seher Abbas and Karl-Wilhelm Koch日期:04/05/2020,浏览量:1091,Q&A: 0
    [Abstract] Diverse and complex molecular recognitions are central elements of signal transduction cascades. The strength and nature of these interaction modes can be determined by different experimental approaches. Among those, Isothermal titration calorimetry (ITC) offers certain advantages by providing binding constants and thermodynamic parameters from ...
    Determination of Flavin Potential in Proteins by Xanthine/Xanthine Oxidase Method
    作者:Elena Maklashina and Gary Cecchini日期:04/05/2020,浏览量:1102,Q&A: 0
    [Abstract] This protocol describes a simple xanthine/xanthine oxidase enzymatic equilibration method for determination of the redox potential of a flavin. As an example of the use of this method, we determine the reduction potential of the covalently bound FAD cofactor (Em = -55 mV) in the SdhA flavoprotein subunit of succinate ...
    Estimating Cellular Abundances of Halo-tagged Proteins in Live Mammalian Cells by Flow Cytometry
    利用流式细胞术进行哺乳动物活细胞中Halo 标记蛋白细胞丰度测定
    作者:Claudia Cattoglio, Xavier Darzacq, Robert Tjian and Anders Sejr Hansen日期:02/20/2020,浏览量:1999,Q&A: 0
    [Abstract] Accurate abundance measurements of cellular proteins are required to achieve a quantitative and predictive understanding of any biological process inside the cell. Existing methods to determine absolute protein abundances are labor-intensive and/or require sophisticated experimental and computational infrastructure (e.g., fluorescence ...
    Evaluating Whole Blood Clotting in vitro on Biomaterial Surfaces
    作者:Roberta M Sabino and Ketul C Popat日期:02/05/2020,浏览量:1574,Q&A: 0
    [Abstract] Biomaterial-associated thrombosis is still a major concern for blood-contacting implants. After the medical device is implanted and comes in contact with blood, several complex reactions occur, which may lead to thrombus formation and failure of the device. Therefore, it is essential to evaluate the biomaterial interaction with the whole blood. ...
    Semi-quantitative Determination of Protein Expression Using Immunohistochemistry Staining and Analysis: An Integrated Protocol
    作者:Alexandra R Crowe and Wei Yue日期:12/20/2019,浏览量:2689,Q&A: 0
    [Abstract] Semi-quantitative IHC is a powerful method for investigating protein expression and localization within tissues. The semi-quantitative immunohistochemistry (IHC) involves using software such as free software ImageJ Fiji to conduct deconvolution and downstream analysis. Currently, there is lack of an integrated protocol that includes a detailed ...
    Separation of Natural Collagen Crosslinks Using Buffer and Ion-pairing Agent Free Solvents on Silica Hydride Column for Mass Spectrometry Detection
    作者:Rafea Naffa and Joseph Pesek日期:05/05/2019,浏览量:2034,Q&A: 0
    [Abstract] In this protocol we describe the separation of collagen crosslinks in biological tissues and samples including skin, tendon, cartilage, bone and urine. The existing methods use either cation exchange chromatography followed by post-column derivatization with ninhydrin or reverse phase chromatography with mass spectrometry detection. The cation ...