Mastering Western Blotting: The Complete Procedure for Perfect Results
Автор: AI Deciphered
Загружено: 2026-01-03
Просмотров: 2
Описание:
Looking for a foolproof Western Blotting protocol? In this video, we provide a complete, technical walkthrough of the entire Western Blot process. From initial protein extraction to final signal detection, we cover the "how-to" and the "pro-tips" for each stage to help you get clean, publishable blots.
Detailed Workflow Covered:
Sample Preparation: Lysis buffers, protease inhibitors, and protein quantification (BCA/Bradford).
Gel Electrophoresis: Setting up the SDS-PAGE, loading samples, and running the gel.
The Transfer: Wet vs. Semi-dry transfer methods and checking efficiency with Ponceau S staining.
Blocking: Choosing between Milk and BSA to reduce non-specific binding.
Antibody Incubation: Primary and secondary antibody dilutions and incubation times.
Detection: ECL Chemiluminescence vs. Fluorescence and how to use the imager.
Why watch this? Western Blotting is notorious for "ghost bands" and high background noise. This guide focuses on the technical nuances that ensure high specificity and reproducibility in your experiments.
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