User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/02/18

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Summary

  • Short summary

Materials & Methods

Materials


Method

Coating of plate

  • Mix
    • 24 mL coating buffer
    • 80 μL primary antibody
  • Add 100 μL per well (96-wells plate)
  • Incubate ON

Preparing Sandwich

  • Wash coated plate 5x 200 μL PBS
  • Remove PBS
  • Add 200 μL BSA
  • Incubate 1h @ RT and remove BSA
    • Dilute samples as shown below for each 24-wells (dilution buffer (μL)/sample (μL))


24-wells (dilution buffer (μL)/sample (μL))
1 2 3 4 5 6
A 100/100 100/100 100/100 100/100 100/100 100/100
B 150/50 150/50 150/50 175/25 175/25 175/25
C 150/50 150/50 150/50 175/25 175/25 175/25
D 150/50 150/50 150/50 175/25 175/25 175/25


  • Wash of BSA with 200 μL Washing buffer 5x
  • Add 100 μL of diluted sample
  • Incubate ≥1h @ RT

Notes

  • Of the 96-wells plate well F1 contain a Polystyrene fragment

Results

  • WHAT?

Conclusion

  • DONE Ow..

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