User:Eric Ma/Notebook/MICB323 Lab Book/2009/03/17

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Experimental Goals

  1. Perform an Immunoprecipitation of iNOS/NOS2 from LPS +/- IFNg stimulated cell lysates.
  2. Preparation of 8% PAGE running gel.

Part One: IP of iNOS/NOS2

  • Volumes of RIPA buffer required for IP. (See previous entry for calculation of volumes.)
  • I am student A, Sunny is Student B, therefore the samples are divided as follows:
Experimenter Sample Number Sample Volume of sample required (µL) Volume of RIPA buffer needed (µL)
Eric 1 24 hr (+) LPS 64.9 285.1
Eric 2 48 hr (+) LPS 74.7 275.3
Eric 3 72 hr (+) LPS 97.9 252.1
Eric 4 24 hr (+) LPS (+) IFNg 92.6 257.4
Eric 5 48 hr (+) LPS (+) IFNg 128.8 221.2
Eric 6 72 hr (+) LPS (+) IFNg 62.5 287.5
Sunny 7 24 hr (+) IFNg 34.9 315.1
Sunny 8 48 hr (+) IFNg 40.3 309.7
Sunny 9 72 hr (+) IFNg 350.0 0.0
Sunny 10 24 hr control 34.2 315.8
Sunny 11 48 hr control 35.6 314.4
Sunny 12 72 hr control 34.2 315.8

Part Two: 8% Acrylamide Gel

  • Done with no problems encountered.