UNSW Cell Biology

ANAT3231 Laboratory 04

Introduction

This laboratory is an introduction to methods of cell fractionation for analysis of cells and tissues in cell biology. The laboratory handout and textbook alone contain enough information as an introduction to the subject for this level of study. If you are interested in further reading, I have also included below links to more detailed textbooks and websites with further information and images. Please note this additional information is not necessarily examinable, but may be useful if you have not previously studied biology.

Objectives

Understand historic background of cell fractionation

Understand development of major cell fractionation techniques

Brief understanding of cell fractionation

Brief understanding of protein chemistry separation

Page Links: Start Here | NLM Online Textbooks | Cell Organelle Size and Density | Web Links | References | Related Topics | Commercial Resources

Start Here

This Lab content is covered in some good detail in Molecular Biology of the Cell (III. Methods, 8. Manipulating Proteins, DNA, and RNA) Fractionation of Cells We will be using this section as a guide for the Laboratory.

Cell Organelle Size and Density

Nuclei Diameter (5-10 µm) Density (1.4 g/cm3)

Mitochondria Diameter (1-2 µm) Density (1.1 g/cm3)

Ribosomes Diameter (0.02 µm) Density (1.6 g/cm3)

Lysosomes Diameter (1-2 µm) Density (1.1 g/cm3)

Rough Endoplasmic Reticulum Density (1.2 g/cm3) due to bound ribosomes

Golgi vesicles Density (1.14 g/cm3)

Plasma Membranes Density (1.12 g/cm3)

Source: Carolina Tips, Nov. 1, 1973, and Mol. Cell Biol. 1999.

NLM Online Textbooks

Molecular Biology of the Cell 4

NCBI MBoC | Publisher (Garland) MBoC

Molecular Cell Biology

NCBI MCB | Publisher (Freeman) MCB

Purification of Cells and Their Parts

The Cell- A Molecular Approach

NCBI The Cell | Publisher (Sinauer) The Cell

Subcellular Fractionation

Image: Subcellular Fractionation

Image: Velocity centrifugation in a density gradient

Web Links

External

Commercial Resources
SumanasincCell Fractionation
ChemiconCrude Membrane Preparations
BiocompareCell Fractionation Kits

References

Journals

Journal of Histochemistry and Cytochemistry
Histochemistry and Cell Biology
Biotechnic and Histochemistry
European Journal of Histochemistry
Journal of Molecular Histology

Related Topics

organelle labels, aldehyde cross-linking, artefatcs of fixation, immunochemistry, monoclonal antibodies