Merck
CN
  • Protocol for High-Throughput Screening of Neural Cell or Brain Tissue Protein Using a Dot-Blot Technique with Near-Infrared Imaging.

Protocol for High-Throughput Screening of Neural Cell or Brain Tissue Protein Using a Dot-Blot Technique with Near-Infrared Imaging.

STAR protocols (2020-10-13)
Anna C Chlebowski, Glen E Kisby
ABSTRACT

Dot blotting allows for the rapid screening of a larger number of samples and/or targets than more traditional methods, such as a western blot or in-tissue-based methods. We have developed a dot-blot assay specifically for use with a LiCor Odyssey CLx imager, which allows for sensitive detection of proteins in the infrared range. Here, we provide a detailed protocol for the preparation of brain tissue and neural cell culture lysates for analysis of protein targets by dot blotting.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Magnesium chloride, anhydrous, ≥98%
Sigma-Aldrich
Adenosine 5′-triphosphate disodium salt hydrate, microbial, BioReagent, suitable for cell culture, ≥99% (HPLC)
Sigma-Aldrich
TWEEN® 20, for molecular biology, viscous liquid
Roche
cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail, Protease Inhibitor Cocktail Tablets provided in a glass vial, Tablets provided in a glass vial