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High Throughput, Absolute Determination of the Content of a Selected Protein at Tissue Levels Using Quantitative Dot Blot Analysis (QDB)
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
High Throughput, Absolute Determination of the Content of a Selected Protein at Tissue Levels Using Quantitative Dot Blot Analysis (QDB)

High Throughput, Absolute Determination of the Content of a Selected Protein at Tissue Levels Using Quantitative Dot Blot Analysis (QDB)

DOI:
10.3791/56885-v

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08:10 min

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August 21, 2018

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, , , , , , , ,
1Medicine and Pharmacy Research Center,Binzhou Medical University, 2Yantai Zestern Biotechnique Co. LTD, 3Precision Medicine research center,Binzhou Medical University, 4Department of Chemistry – BMC,Uppsala University

Chapters

  • 00:04Title
  • 01:19Sample Preparation
  • 02:22Defining the Linear Range of the QDB Analysis
  • 03:19Process of QDB Analysis
  • 06:11Results: Representative Result of a QDB Analysis to Determine the Absolute CAPG levels in Three Mouse Tissues
  • 06:56Conclusion

Summary

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Here we demonstrate a detailed process of quantitative dot blot analysis (QDB) by determining the absolute content of a targeted protein, capping actin protein, gelsolin-like (CAPG), in three different mouse tissues. We demonstrate a high throughput, convenient, quantitative immunoblot technique for biomarker validation at the cellular and tissue level.

Tags

Quantitative Dot Blot AnalysisQDBHigh-throughputQuantitativeAbsoluteProtein QuantificationProteomicsBiomarker VerificationTriton X Lysis BufferBCA AssayRecombinant Protein Standard

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