User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/06/21

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Summary

  • 12June2010: CSE & S0 (hTERT D9 P27)
  • 15June2010: CSE & S0 (hTERT D9 P28)
  • 17June2010: CSE & S0 (hTERT D9 P29)
  • Got some new HEK293 cells, in about 7.5 mL (2·106 cells/mL) Medium
    • Spinned down 1000 rcf 5 min.
      • NOTE: First 4000 rcf for a few seconds! (maybe not nice for cells, but needed to lyse them anyway)
    • Resuspended the cells twice in cold PBS (non-sterile) followed by spinning down (1000 rcf, 5 min.) and removing supernatant
    • Added 2 mL of RIPA lysis buffer and sonicated them

Bradford assay

  • Dilute samples 5x (4 μL samples + 16 μL H2O)
  • Put (3x) 5 μL sample on plate
  • Put STND on plate
  • Add 250 μL Bradford reagent
  • Measure (AG KLUSSMANN FRANK BRADFORD ASSAY)

Materials & Methods

Materials

Method

6-wells
# 1 2 3
A
B
24-wells
# 1 2 3 4 5 6
A
B
C
D
96-wells plate
# 01 02 03 04 05 06 07 08 09 10 11 12
A
B
C
D
E
F
G
H
I

Notes

Results

  • WHAT?

Discussion

  • DONE Ow..

Related

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Attachment

<html> <a href="http://openwetware.org/wiki/Image:Bradford_21062010.xls"> 

  <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50">

</a> </html> Bradford results


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