Cell and tissue lysis hub

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(comparison of lysis method section)
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* [[Jacobs:Protocol Total Protein Isolation Using RIPA Lysis Buffer]] - Jacobs Lab: cell line lysis for protein (method: RIPA buffer)
* [[Jacobs:Protocol Total Protein Isolation Using RIPA Lysis Buffer]] - Jacobs Lab: cell line lysis for protein (method: RIPA buffer)
* [[Streptomyces:Protocols/Mini-Maxi Prep]] - Streptomyces community, East Anglia: bacteria lysis for DNA (method: NaOH)
* [[Streptomyces:Protocols/Mini-Maxi Prep]] - Streptomyces community, East Anglia: bacteria lysis for DNA (method: NaOH)
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== Comparison of lysis methods ==
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=== Sonication ===
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* most efficient method of cell fractionation
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* problem: heat build up which can denature proteins (proportionate to length of sonication)
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* precaution: do on ice and sonicate intermittantly
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=== Homogenisation ===
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* best for animal tissue; less suitable for cells
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* precaution: do on ice to reduce heat build-up and denaturation
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=== Freeze-thaw ===
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* least effective method
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* plus: does not denature proteins as much as other methods
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=== Detergents ===
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* chemical method of lysis
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* problems: detergent may inhibit subsequent reactions
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* problems: detergent may disrupts protein interactions

Revision as of 05:16, 26 June 2008

This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison.


Contents

Comparison of lysis methods

Sonication

  • most efficient method of cell fractionation
  • problem: heat build up which can denature proteins (proportionate to length of sonication)
  • precaution: do on ice and sonicate intermittantly

Homogenisation

  • best for animal tissue; less suitable for cells
  • precaution: do on ice to reduce heat build-up and denaturation

Freeze-thaw

  • least effective method
  • plus: does not denature proteins as much as other methods

Detergents

  • chemical method of lysis
  • problems: detergent may inhibit subsequent reactions
  • problems: detergent may disrupts protein interactions
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