Titlebook: High-Resolution Imaging of Cellular Proteins; Methods and Protocol Steven D. Schwartzbach,Omar Skalli,Thomas Schikors Book 2016 Springer Sc

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Immunoelectron Microscopy of Cryofixed Freeze-Substituted Yeast ,g offers an excellent way to fix cellular structure. However, its use for immunolabeling has remained limited because of the low frequency of labeling due to loss of protein antigenicity or accessibility. Here we present a protocol for immunogold labeling of the yeast . that gives specific and multi

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Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Ug systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. With these methodologies, glycan structures and biological functions are being elucidated. In the study of glycan function as well as disease diagnosis, it is important to e

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Production and Purification of Polyclonal Antibodieslls in response to an immunogen. Polyclonal antibodies raised against an antigen recognize multiple epitopes on a target molecule, which results in a signal amplification in indirect immunoassays including immune-electron microscopy. In this chapter, we present a basic procedure to generate polyclonal antibodies in rabbits.

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Preparation of Colloidal Gold Particles and Conjugation to Protein A/G/L, IgG, F(ab′)2, and Streptavells and tissues by immunoelectron microscopy (IEM). This chapter describes different methods for the preparation of colloidal gold and conjugation of colloidal gold to protein A/G/L, IgG, and streptavidin.

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Bimolecular Fluorescence Complementation (BiFC) Analysis of Protein–Protein Interactions and Assessmn example to provide a step-by-step BiFC protocol using a Nikon A1 confocal microscope and NIS-Elements imaging software. The protocol given below can be readily adapted for use with other confocal microscopes or imaging software.