Klapperich Lab:Photografting to Zeonor Slides

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Overview

This protocol describes the procedure for grafting PEG-acrylate to Zeonor slides.

Materials

  • PEG acrylate
  • Ethylene Glycol
  • 2-Dimethoxy-2-acetophenone (DMPAP)
  • Zeonor slides
  • methanol

Procedure

  1. Weigh out 35 mg DMPAP
  2. Clean plastic slide with EtOH, then rinse with Millipore water
  3. Plasma treat surface of plastic slide for 2 min on medium
  4. Mix DMPAP with 800Failed to parse (MathML with SVG or PNG fallback (recommended for modern browsers and accessibility tools): Invalid response ("Math extension cannot connect to Restbase.") from server "https://api.formulasearchengine.com/v1/":): {\displaystyle u} L Ethylene Glycol and 1200Failed to parse (MathML with SVG or PNG fallback (recommended for modern browsers and accessibility tools): Invalid response ("Math extension cannot connect to Restbase.") from server "https://api.formulasearchengine.com/v1/":): {\displaystyle u} L PEG-acrylate in 2mL tube to get 60% PEG-acrylate solution
  5. Heat solution in hot water bath at 85&degC until DMPAP crystals have melted
  6. Vortex solution
  7. Add solution in uniform layer plastic slide
  8. Photograft slide in UV Crosslinker for 30 seconds on maximum energy
  9. Rinse with MeOH and let dry under hood
  10. remove excess poly-PEG acrylate

Notes

  1. List troubleshooting tips here.
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References

Relevant papers and books

  1. Goldbeter A and Koshland DE Jr. An amplified sensitivity arising from covalent modification in biological systems. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6840-4. PubMed ID:6947258 | HubMed [Goldbeter-PNAS-1981]
  2. JACOB F and MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318-56. PubMed ID:13718526 | HubMed [Jacob-JMB-1961]
  3. ISBN:0879697164 [Ptashne-Genetic-Switch]
All Medline abstracts: PubMed | HubMed

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