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Quantitative Immunoblotting of Cell Lines as a Standard to Validate Immunofluorescence for Quantifying Biomarker Proteins in Routine Tissue Samples
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Quantitative Immunoblotting of Cell Lines as a Standard to Validate Immunofluorescence for Quantifying Biomarker Proteins in Routine Tissue Samples

Quantitative Immunoblotting of Cell Lines as a Standard to Validate Immunofluorescence for Quantifying Biomarker Proteins in Routine Tissue Samples

DOI:
10.3791/58735-v

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09:58 min

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January 07, 2019

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, , ,
1Department of Pathology and Molecular Medicine,Queen’s University, 2Division of Cancer Biology and Genetics,Queen’s Cancer Research Institute, 3Queen’s Laboratory for Molecular Pathology, Department of Pathology and Molecular Medicine,Queen’s University

Chapters

  • 00:04Title
  • 01:29Building a Cell-Line Tissue Microarray (TMA)
  • 03:37Sample Staining by Immunofluorescence
  • 04:27Quantitative Immunoblotting of Cell Lines
  • 07:48Results: Immunoblotting Ratios Strongly and Positively Correlated with Intensity Readings from Quantitative IF
  • 09:22Conclusion

Summary

Automatic Translation

Automatic Translation

We describe the use of quantitative immunoblotting to validate immunofluorescence histology coupled with image analysis as a means of quantifying a protein of interest in formalin-fixed, paraffin-embedded (FFPE) tissue samples. Our results demonstrate the utility of immunofluorescence histology for ascertaining the relative quantity of biomarker proteins in routine biopsy samples.

Tags

Keyword Extraction:Quantitative ImmunoblottingCell LinesImmunofluorescenceBiomarker ProteinsCancer PathologyImmunohistochemistryProtein QuantificationImmortalized Cell LinesMultiplexed ApproachPrimary Biopsy SamplesCentrifugationCell PelletPBSNBFAgarose Solution

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