Shreffler:Notebook/Alex Daily/2012/03/21

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SDS-PAGE on peanut protein from Mt Sinai

Ran SDS-PAGE with Bert to gauge protein purity/concentration of Ara h 2, Ara h 3, and Ara h 6 stock from Mt Sinai, each with a stated concentration of 3.4 mg/ml. Ran nanodrop to determine approximate protein concentrations:

  • Ara h 2: ~1 mg/ml
  • Ara h 3: ~2 mg/ml
  • Ara h 6: ~0.5 mg/ml

Made loading solutions for each protein as follows:

  • Arah h 2: 11 µl protein stock, 2 µl PBS, 5 µl SB, 2 µl RA
  • Arah h 3: 5 µl protein stock, 8 µl PBS, 5 µl SB, 2 µl RA
  • Arah h 6: 13 µl protein stock, 5 µl SB, 2 µl RA

Loaded the gel w/ 1 µg and 5 µg for each protein.

20120320 Gel Arah2 3 6.jpg

According to Bert, band sizes, when matched to bands of known protein concentration (from his past gels), reflected nanodrop concentrations fairly well.