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

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==Part One: IP of iNOS/NOS2==
==Part One: IP of iNOS/NOS2==
* Volumes of RIPA buffer required for IP.
* Volumes of RIPA buffer required for IP. (See previous entry for calculation of volumes.)
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| align="center" style="background:#f0f0f0;"|'''Sample'''
| align="center" style="background:#f0f0f0;"|'''Sample'''

Revision as of 10:23, 17 March 2009

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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.)
Sample [Protein] (mg/mL) Amount of protein in 350 µL (mg) Volume needed to equal 0.042 mg (µL) Volume of RIPA buffer needed (µL)
24 hr (+) LPS 0.651 0.228 64.9 285.1
48 hr (+) LPS 0.566 0.198 74.7 275.3
72 hr (+) LPS 0.432 0.151 97.9 252.1
24 hr (+) IFNg 1.212 0.424 34.9 315.1
48 hr (+) IFNg 1.048 0.367 40.3 309.7
72 hr (+) IFNg 0.121 0.042 350.0 0.0
24 hr (+) LPS (+) IFNg 0.456 0.160 92.6 257.4
48 hr (+) LPS (+) IFNg 0.328 0.115 128.8 221.2
72 hr (+) LPS (+) IFNg 0.676 0.237 62.5 287.5
24 hr control 1.237 0.433 34.2 315.8
48 hr control 1.188 0.416 35.6 314.4
72 hr control 1.237 0.433 34.2 315.8

Part Two: 8% Acrylamide Gel


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