User:Linh N Le/Notebook/2009/06/23: Difference between revisions

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***5 mM EGTA, pH 7.0  
***5 mM EGTA, pH 7.0  
***2 mM MgCl2 ( final Concentration: 2 mM)
***2 mM MgCl2 ( final Concentration: 2 mM)
 
*[http://www.ece.cmu.edu/~yuliwang/Methods/Materials/Fluorescent%20Labelilng/RhodamineTubulin.pdf Yu-li Wang from Carnegie Mellon] This is a pdf of how they do Rhodamine Tubulin
** PEM buffer:
***0.1 M PIPES
***1.0 mM EGTA
***0.5 mM MgCl2
***pH 6.9




Revision as of 14:09, 23 June 2009

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To Do

  • Koch/Atlas group meeting at 3
  • Try out new buffer we made yesterday?
  • Try Kinesin?
  • Clean up lab

BRB80

  • Since there are so many ways to make this buffer, Koch wanted me to try to find different famous people's recipes for the buffer
    • People to look for, Block and Vale

Recipes

  • Hancock (Used by us as well)
    • 80 mM PIPES pH 6.85 (we use ph6.9)
    • 1 mM MgCl2
    • 1 mM EGTA
  • T.Salmon Labs
    • PM Buffer
      • 100 mM PIPES, pH 6.9
      • 2 mM EGTA
      • 1 mM Mg2SO4
  • Goldman Lab (Northwestern University)
    • 100 mM Pipes,pH 6.9
    • 1 mM MgCl2
    • 1 mM EGTA
  • Cold Spring Harbor Labs
    • PEM Buffer
      • 0.1 M PIPES (pH 6.95)
      • 2 mM EGTA
      • 1 mM MgSO4
  • Cytoskeleton
    • PEM (General Tubulin Buffer)
      • 80 mM Na-PIPES pH 6.9
      • 1 mM MgCl2
      • 1 mM EGTA
  • Mitchison Lab (Harvard)
    • Brinkley Buffer 1980 (BRB80)
      • 80mM PIPES pH 6.8
      • 1mM MgCl2
      • 1mM EGTA
    • "Cytoskeleton Buffer"
      • 10mM MES pH 6.1
      • 138mM KCl
      • 3mM MgCl
      • 2mM EGTA
    • Notes: BRB80 is good for microtubules and 'cytoskeleton buffer' is good for both actin filaments and microtubules
  • Sigma Aldrich
    • PEM Buffer:
      • 0.1M PIPES (Product No. P8203)
      • 5 mM EGTA (Product No. E4378)
      • 2mM MgCl2 · 6H2O (Product No. M0250)
      • Bring to pH 6.8, using NaOH solution
  • Bayer College of Medicine
    • PEM Buffer:
      • 80mMPotassium PIPES, pH 6.8
      • 5 mM EGTA, pH 7.0
      • 2 mM MgCl2 ( final Concentration: 2 mM)
  • Yu-li Wang from Carnegie Mellon This is a pdf of how they do Rhodamine Tubulin
    • PEM buffer:
      • 0.1 M PIPES
      • 1.0 mM EGTA
      • 0.5 mM MgCl2
      • pH 6.9


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