User:Eric Ma/Notebook/MICB323 Lab Book/2009/03/24: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
Line 38: Line 38:


==Part 2: SDS-PAGE==
==Part 2: SDS-PAGE==
Wells loaded as follows.
* Wells loaded as follows.


* Notes:
* Notes:

Revision as of 15:06, 24 March 2009

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Goals

  1. Perform ELISA to detect TNFa in LPS +/- IFNg stimulated culture supernatants.
  2. Perform PAGE on the IP samples from Lab 8
  3. Western Transfer following PAGE


Part 1: ELISA Detection of TNFa in LPS +/- IFNg stimulated culture supernatants.

  • 96 well immunoplate layout:
Row Row 1-4 Row 5-8 Row 9-12
A 0 pg TNFa (assay buffer only) 0 hr PBS control Day 1 +PBS control
B 62.5 pg/mL TNFa 30 min +LPS 30 min +LPS +IFNg
C 125 pg/mL TNFa 60 min +LPS 60 min +LPS +IFNg
D 250 pg/mL TNFa 90 min +LPS 90 min +LPS +IFNg
E 500 pg/mL TNFa Supplied 6 hr +LPS sample Supplied 6 hr +LPS +IFNg
F 1000 pg/mL TNFa Supplied 8 hr +LPS sample Supplied 8 hr +LPS +IFNg
G 2000 pg/mL TNFa 1/5 dilution of Day 1 +LPS stimulation 1/5 dilution Day 1 +LPS +IFNg
H 4000 pg/mL TNFa 90 min + IFNg Day 1 +IFNg

Part 2: SDS-PAGE

  • Wells loaded as follows.
  • Notes:
    • The gel ran in a funny manner - it was uneven, and resulted in slanting. As the pattern was identical on both my plate and Sunny's plate (i.e. ours were mirror images of each other), we think it may be due to a voltage problem. The wiring may be causing this.
    • As a result, it will be hard for us to measure any sizes of the proteins.


Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Eric_Ma/Notebook/MICB323_Lab_Book/2009/03/24&oldid=296486"