Applied Biomics offers improved 2D Western blot

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  • Published: Oct 5, 2016
  • Author: Jon Evans
  • Source: Applied Biomics
  • Channels: Electrophoresis

The US proteomics company Applied Biomics is offering a novel fluorescent 2D Western blot (WB) with significant advantages and superior data quality over traditional WBs.

The traditional 2D WB requires running two 2D gels with the same sample: one gel is silver stained and the other gel is used for WB. The reactive protein spots are determined by comparing the stained gel to the WB gel, but this procedure suffers from gel-to-gel variations. When calculating the antibody coverage, one assumes that all the proteins are completely transferred, which is not always the case. In addition, silver staining is not a quantitative method.

Applied Biomics' fluorescent 2D WB overcomes these limitations and offers significant advantages.

  1. High consistency: Fluorescent 2D Western detects both protein and WB images on the same gel, allowing perfect alignment between protein and WB spots, thus eliminating any gel-to-gel variations.
  2. Cost-effective: Since only one gel is needed, it saves the cost of running duplicate gels.
  3. High accuracy: Fluorescent CyDye labeling allows accurate quantitation of protein and WB spots.
  4. High sensitivity: Fluorescent CyDye labeling has a higher sensitivity and wider dynamic range than silver staining.

Together with its proprietary sample preparation protocols, Applied Biomics has successfully used its fluorescent 2D WB for various applications.

  1. Characterizing antibody coverage of host cell protein (HCP) antibodies.
  2. Detecting HCP contaminants originating from cells used to manufacture therapeutic proteins and vaccines.
  3. Identifying important biomarkers.

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