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

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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)