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Whilst Western blotting remains the pre-eminent method of choice for protein detection and characterisation, the various improvements to individual aspects of Western work flow developed in recent years have had little impact on the relatively poor reproducibility, lack of accurate quantification, extensive time to result and reliability issues associated with traditional Western blotting.

Wes employs capillary electrophoresis combined with chemi-luminescence based antibody detection to run the entire Western immunoassay process from start to finish, providing dramatic improvements in sensitivity, reproducibility, quantitative accuracy and time to result compared to all other legacy Western blotting methods. Simple Western assays cover three separation ranges: 2 – 40kDa, 12 – 230 kDa and 66 – 440kDa.

Wes delivers exceptional results for virtually all sample types requiring a Western blotting approach. Minimal sample volume (3-5uL) at a low total protein concentration (0.2ug/uL) are required. Runs typically comprise 24 samples plus a size ladder and full results are generated in just over 3 hours. The Simple Western assay uses quality controlled proprietary consumables and all the client would ordinarily need to supply are their samples and a suitable primary antibody. An ever expanding wide range of Simple Western certified primary antibodies are available and primary antibodies from other sources may also be used.

Application of WES: Western Assay System:

Autophagy assay: LC-3 is a protein that is implicated in the control of autophagy (destructive mechanism of the cell that disassembles unnecessary or dysfunctional components). We used an LC-3 antibody to detect levels of LC-3 in cells following treatment with compounds that both stimulate and inhibit.

 

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