WESTERN BLOTTING AND WESTERN BLOTTING AND B B RD RD U STAINING U STAINING CHRISTIAN LAPITAN CHRISTIAN LAPITAN MILLIKAN HIGH SCHOOL – LONG BEACH AMRITA AHLUWALIA, PH.D. AMRITA AHLUWALIA, PH.D. GASTROINTESTINAL (GI) RESEARCH Amrita Ahluwalia, Ph.D. Gastrointestinal(GI) Research
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
WESTERN BLOTTING WESTERN BLOTTING AND BAND BRDRDU STAININGU STAINING
CHRISTIAN LAPITANCHRISTIAN LAPITANMILLIKAN HIGH SCHOOL – LONG BEACH
AMRITA AHLUWALIA, PH.D.AMRITA AHLUWALIA, PH.D.GASTROINTESTINAL (GI) RESEARCH
Amrita Ahluwalia, Ph.D.Gastrointestinal(GI) Research
Methods learned so far
Cell CultureProtein isolationBradford’s assay for
determining protein concentration
SDS-PAGEWestern TransferImmunoblottingBrdU assay
Cells used:• HCT116 (colon cancer
cells) • NCM460 (normal
colonic mucosal cells)
Western BlotWestern Blot
Protein detection Gel Electrophoresis
Gel matrix: pore size is inversely proportional to gel concentration
Protein samples (-) in the wells move to the positive anode (+)
The size and weight of each protein determines how far it moves Top
Bottom
Amrita Ahluwalia, Ph.D.Gastrointestinal(GI) Research