sds page and western blot protocol · 1 sds page and western blot protocol laboratory - day 1...
Post on 28-Sep-2018
226 Views
Preview:
TRANSCRIPT
1
SDS PAGE and Western BlotProtocol
Laboratory - Day 1
• Bring in fish samples.• Analyze relatedness among samples using
the phylogenic charts.• Complete Laboratory Day 1 protocol
– Laemmli sample buffer• Solubilize the proteins in the fish muscle samples• Tris buffer, SDS, bromophenol blue, and glycerol.
– Molecular weight marker– Actin and myosin standard
2
Lab - Day 1: Extract and Denature FishProteins
Lab Day 2: Gel loading, running andstaining
• Defrost the protein extracts.• Prepare the electrophoresis chamber• Prepare the pre-cast gel and lock it into the
electrophoresis chamber.• Load and run the gel.
– 1x TGS buffer Tris-glycine-SDS• Stain the gel
– Coomassie stain
4
Lab Day 2: Gel loading, running andstaining
Lab Day 2: Gel loading, running andstaining
**Change volume to 5ul for all samples
8
LAB DAY 4: Immunodetectionfor Myosin Light Chains
LAB DAY 4: Immunodetectionfor Myosin Light Chains
top related