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

Revision as of 16:14, 23 June 2009

Project name

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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
What we use for Cytoskeleton is called PEM and while PEM and BRB80 can be the same, they do not have to be
- I did searches on both PEM and BRB80 and the following recipes are also reported by what they are called
Special Note: All PDF's linked here are from the personal website of the authors and are free access

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
  - 100 mM 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
Tedeschi of Italy
- PEM buffer
  - 85 mM Pipes, pH 6.94
  - 10 mM EGTA
  - 1 mM MgCl2
Steve Gross of U Minnesota with Steve Block
- 80 mM Pipes
- 1 mM EGTA
- 4 mM MgCl2
Stefan Diez (Germany)
- BRB80 buffer
  - 80 mM potassium Pipes
  - 1 mM EGTA
  - 1 mM MgCl2
  - pH 6.9
Mathew Lang, Steve Block first author
Also used in Rosenfield, Block Paper
Also used in Guydosh Block Paper with 10μM of Taxol
- Un-named buffer
  - 80 mM Pipes (pH6.9)
  - 50 mM potassium acetate
  - 4 mM MgCl2
  - 2 mM DTT
  - 1 mM EGTA
  - 7 mMTaxol
Valentine, Fordyce, Block
- polymerization buffer for brain tubulin
  - 62 mM PIPES at pH 6.9
  - 0.8 mM EGTA
  - 3.7 mM MgCl2
  - 0.9 mM GTP
  - 10% v/v DMSO
- stablilzation buffer
  - 80 mM PIPES at pH 6.9
  - 1.7 mM EGTA
  - 5.5 mM MgCl2
  - 1.2 mM GTP
  - 8.2 mM sodium azide
  - 20 μM taxol
  - 10% v/v DMSO)

@@ Line 102: / Line 102: @@
 ***0.9 mM GTP
 ***10% v/v DMSO
+**stablilzation buffer
+***80 mM PIPES at pH 6.9
+***1.7 mM EGTA
+***5.5 mM MgCl2
+***1.2 mM GTP
+***8.2 mM sodium azide
+***20 μM taxol
+***10% v/v DMSO)
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User:Linh N Le/Notebook/2009/06/23: Difference between revisions

Revision as of 16:14, 23 June 2009

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