User:David K. Barclay/Notebook/Controlling Pancreas Cell Fate Using Transcription Factors/2014/11/17: Difference between revisions

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* Continuing Protocol from NAP blocking stage. Protocol [http://openwetware.org/wiki/Haynes:ELISA here]
* Continuing Protocol from NAP blocking stage. Protocol [http://openwetware.org/wiki/Haynes:ELISA here]
* Primary Antibodies:
* Primary Antibodies:
      *FL180 (Santa Cruz Biotech)- Glucagon (concentration 200ug/mL) - diluted 1/50
      *FL180 (Santa Cruz Biotech)- Glucagon (Rabbit- concentration 200ug/mL) - diluted 1/50
<br> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp; *ab7842 (ABCAM) - Insulin (concentration 200ug/mL) - diluted 1/50
<br> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp; *ab7842 (ABCAM) - Insulin (Guinea Pig- concentration 200ug/mL) - diluted 1/50
*Insulin primary not meant for ELISA? Testing anyway
*Insulin primary not meant for ELISA? Testing anyway
<br>
* Secondary Antibodies:
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; *sc-2030 (Santa Cruz Biotech)- anti-Rabbit IgG-HRP (200 ug/0.5mL) - diluted 1/500
<br> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp; *sc-2438 (Santa Cruz Biotech - anti-Guinea pig IgG-HRP (200ug/0.5mL) - diluted 1/500


==Pancreatic Cell Culture==
==Pancreatic Cell Culture==

Revision as of 13:06, 17 November 2014

Cell Fate Switch by Synthetic Transcription Factors <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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ELISA (test)

  • Continuing Protocol from NAP blocking stage. Protocol here
  • Primary Antibodies:

      *FL180 (Santa Cruz Biotech)- Glucagon (Rabbit- concentration 200ug/mL) - diluted 1/50
      *ab7842 (ABCAM) - Insulin (Guinea Pig- concentration 200ug/mL) - diluted 1/50

  • Insulin primary not meant for ELISA? Testing anyway


  • Secondary Antibodies:

      *sc-2030 (Santa Cruz Biotech)- anti-Rabbit IgG-HRP (200 ug/0.5mL) - diluted 1/500
      *sc-2438 (Santa Cruz Biotech - anti-Guinea pig IgG-HRP (200ug/0.5mL) - diluted 1/500

Pancreatic Cell Culture

  • Cells look near 60% confluence - FBS worked?

RT-PCR

  • Dilutions of Target Primers to 1/10 (100uM)




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