User:Karmella Haynes/Notebook/PcTF Genomics/2015/03/20

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03/20/15

  • PAGE - glucagon & insulin (for Gbox demo next week)



PAGE - glucagon & insulin
> From David Barclay's pancreas cell project


Sample prep

  • Loading vol. 20 uL = 5.0 μL 4x Novagen sample buffer (blue) + 1.0 μL 1M DTT + 14 uL protein
  • Dilutions - make enough for at least two lanes plus pipetting error (x3)
Protein Final mass Stock μL (x3) dH2O μL (x3)
Glucagon 10.0 μg 10.0 (30.0) 4.0 (12.0)
Glucagon 1.0 μg 1.0 (3.0) 13.0 (39.0)
Glucagon 0.1 μg 0.1 (0.3) 13.9 (41.7)
Insulin* 10.0 μg 10.0 (30.0) 4.0 (12.0)
Insulin* 1.0 μg 1.0 (3.0) 13.0 (39.0)
Insulin* 0.1 μg 0.1 (0.3) 13.9 (41.7)
  • For insulin, start with a 1/25 dilution of the 25 mg/mL stock
  • 4x sample buffer batch (x15) = 75.0 4x sample buffer + 15.0 1M DTT


> Gel lanes (for both gels)

  1. Page Ruler plus (15 uL)
  2. Glucagon 10.0 μg
  3. Glucagon 1.0 μg
  4. Glucagon 0.1 μg
  5. Insulin 10.0 μg
  6. Insulin 1.0 μg
  7. Insulin 0.1 μg
  8. Page Ruler plus (15 uL)
  9. Glucagon 10.0 μg
  10. Glucagon 1.0 μg
  11. Insulin 10.0 μg
  12. Insulin 1.0 μg


Reagent Volume
reagent 1 # μL
reagent 2 #
reagent 3 #
reagent 4 #
dH2O #
  # μL

--> Reaction conditions



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