Conceptual
Browse Instruct Curate
Login

About Western Blotting: From Lysate to Quantified Band

Copied

You will learn how to take a biological sample from lysis through separation, transfer, immunodetection, and quantification without producing an uninterpretable blot. You will be able to choose a lysis buffer and inhibitor set for your target, load equal protein by a validated assay, pick a gel percentage and transfer method matched to your protein's mass, and verify transfer before you spend antibody. You will also be able to titrate antibodies, distinguish chemiluminescent from fluorescent detection and their dynamic-range limits, normalize to total protein rather than a single housekeeping band, and diagnose the common failure modes from the appearance of the blot.

Estimated Time to Complete

Only available after login

What You'll Learn

Concepts:
Enzyme-Conjugated Secondary Antibody Enhanced Chemiluminescence Detection Total Protein Normalization Western Blot Artifact Diagnosis SDS-PAGE Protein Separation Electrophoretic Protein Transfer Membrane Stripping and Reprobing Antibody Validation Cell Lysis Buffer Selection Fluorescent Western Blot Detection BCA Protein Assay Primary Antibody Titration Signal Saturation and Dynamic Range Membrane Blocking Protease and Phosphatase Inhibition Densitometric Band Quantification

What you will learn

No introduction video available

About Demerzel

D

Guide profile coming soon.