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Using total protein lysates for Western blotting confers greater benefits than traditional approaches that use fractionated samples, where a portion of the sample is discarded during processing. By incorporating low-volume inputs, it is possible to identify any protein present in the sample where a suitable antibody is available. The inclusion of a calibration curve allows quantitative analyses to be undertaken. There are additional benefits including that only requiring a small input saves on precious samples. Using whole muscle lysates is necessary in order to ascertain specificity of antibodies for subsequent immunofluorescent methods.

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Extraction of Total Protein from Cardiomyocytes and Western Blotting Analysis

  • 实验方案

  • 发表日期：

    2025年

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  • James A. Loehr,
  • Heidy K. Latchman,
  • Robyn M. Murphy,
  • Tanya R. Cully

摘要

Using total protein lysates for Western blotting confers greater benefits than traditional approaches that use fractionated samples, where a portion of the sample is discarded during processing. By incorporating low-volume inputs, it is possible to identify any protein present in the sample where a suitable antibody is available. The inclusion of a calibration curve allows quantitative analyses to be undertaken. There are additional benefits including that only requiring a small input saves on precious samples. Using whole muscle lysates is necessary in order to ascertain specificity of antibodies for subsequent immunofluorescent methods.