Laboratory Methods in Enzymology: Cell, Lipid and Carbohydrate

Series Editor:

  • Jon Lorsch, Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA

Methods in Enzymology volumes provide an indispensable tool for the researcher. Each volume is carefully written and edited by experts to contain state-of-the-art reviews and step-by-step protocols.

In this volume, we have brought together a number of core protocols concentrating on Cell, Lipid and Carbohydrate, complementing the traditional content that is found in past, present and future Methods in Enzymology volumes.

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Audience

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

 

Book information

  • Published: November 2013
  • Imprint: ACADEMIC PRESS
  • ISBN: 978-0-12-420067-8


Table of Contents

Pouring Agar Plates and Streaking or Spreading to Isolate Individual Colonies

Storage of Bacteria and Yeast

Cellular Fractionation - Mammalian Cells

Cellular Fractionation - Yeast cells

Yeast-Gene Replacement Using PCR Products

Measuring RNAi knockdown using qPCR

Recombineering: Using Drug Cassettes to Knock out Genes in vivo

Gene Knockouts, in vivo Site-Directed Mutagenesis and Other Modifications Using the Delitto Perfetto System in Saccharomyces cerevisiae

Gene targeting and Site-specific recombination in Mouse ES Cells

Recombineering: Highly Efficient in vivo genetic engineering using single-strand oligos

Growth Media for E. coli

Saccharomyces cerevisiae Growth Media

Immunohistochemistry on Freely Floating Fixed Tissue Sections

Preparation of Slides for Microscopy from Frozen Tissue Sections

Preparation of Formalin-fixed Paraffin-embedded Tissue for Immunohistochemistry

Preparation of Cells for Microscopy using Cytospin

Preparation of Cells for Microscopy using Chamber Slides and Coverslips

Preparation of Cells for Microscopy using ‘Cell Blocks’

Enzymatic deglycosylation of glycoproteins

Preparation of fatty acid or phospholipid vesicles by thin-film rehydration

Vesicle extrusion through polycarbonate track-etched membranes using a hand-held mini-extruder

Preparation of fatty acid micelles

Reverse-phase HPLC Analysis and Purification of Small Molecules

Thin Layer Chromatography

ATP and GTP hydrolysis assays (TLC)