8F2B0021-C34B-4D43-AF91-CE203C00F994.jpeg - ckspace NG 3:50 PM eros\LI 3 Diff DHFR protein Lab 5 DHFR Fusion proteins for analysis tags His GST Myc Flag

8F2B0021-C34B-4D43-AF91-CE203C00F994.jpeg - ckspace NG 3:50...

This preview shows page 1 out of 1 page.

Image of page 1

You've reached the end of your free preview.

Want to read the whole page?

Unformatted text preview: ckspace NG 3:50 PM 1/16/2019 eros\ \LI 3 Diff DHFR protein Lab 5: DHFR Fusion proteins for analysis tags! - His - GST - Myc Flag mamallen You will be analyzing three DHER fusion proteins, each is a fusion of human DHFR with a different wheat punned . Purified it Stagged DHFR, expressed and purified from E. coli germ . Purified GST -tagged DHFR, in vitro-translated from wheat germ extract . Whole cell lysate containingMy Flag cell - free) mammalian cell line tagged DHFR, from expression in a Ecoli His-tag My cflag Puired proten tag GSETtag Transfer get to pitter paper to look for Western Blot analysis of fusion proteins specific protein . For Western Blot: Gwhen we do a blot . Transfer proteins from an SDS-PAGE gel to a nitrocellulose membrane and then analyze proteins on the membrane. We are looking for one Allows us to look for SPECIFIC bands belonging to a protein of interest SDS-PAGE?? re to Coomassie/Gel code blue analysis by Protein ( doesn't show . Use Western Blotting to EXPERIMENTALLY find the weight- of your tagged DHFR proteins . Then compare these values to the ones computed via sequence analysis in part 5d au proteins ) . Gel given for transfer was SDS-Page gel 47 transfer to [email protected] Recap: SDS: negatively charged detergent specific proteins ( antibode bME: reducing agent HEATED WITH SOS AND MERCAPTOETHANKS don't transfer on gel )...
View Full Document

  • Fall '07
  • Sikorski

What students are saying

  • Left Quote Icon

    As a current student on this bumpy collegiate pathway, I stumbled upon Course Hero, where I can find study resources for nearly all my courses, get online help from tutors 24/7, and even share my old projects, papers, and lecture notes with other students.

    Student Picture

    Kiran Temple University Fox School of Business ‘17, Course Hero Intern

  • Left Quote Icon

    I cannot even describe how much Course Hero helped me this summer. It’s truly become something I can always rely on and help me. In the end, I was not only able to survive summer classes, but I was able to thrive thanks to Course Hero.

    Student Picture

    Dana University of Pennsylvania ‘17, Course Hero Intern

  • Left Quote Icon

    The ability to access any university’s resources through Course Hero proved invaluable in my case. I was behind on Tulane coursework and actually used UCLA’s materials to help me move forward and get everything together on time.

    Student Picture

    Jill Tulane University ‘16, Course Hero Intern

Ask Expert Tutors You can ask You can ask ( soon) You can ask (will expire )
Answers in as fast as 15 minutes