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An In Vitro Organ Culture Model of the Murine Intervertebral Disc
Published on: April 11, 2017
Immunohistochemical analysis of protein expression in formalin fixed paraffin embedded human intervertebral disc
Abbie Binch1, Joseph Snuggs1, Christine L Le Maitre1
1Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK.
Insights
Immunohistochemistry (IHC) is crucial for detecting protein expression in adult human intervertebral disc (IVD) tissues, overcoming autofluorescence issues that hinder other methods. This guide provides optimized IHC protocols for formalin-fixed, paraffin-embedded IVD tissues.
Area of Science:
- Biochemistry
- Histology
- Immunology
Background:
- Adult human intervertebral disc (IVD) tissues exhibit significant autofluorescence, complicating immunofluorescence techniques.
- Immunohistochemistry (IHC) is a preferred method for protein localization and semiquantification in IVD tissues due to its robustness against autofluorescence.
- IHC relies on specific epitope-antibody interactions to detect antigen expression within tissue samples.
Purpose of the Study:
- To discuss the principles of IHC as applied to formalin-fixed paraffin-embedded (FFPE) adult human IVD tissue.
- To provide optimized IHC protocols for antibodies commonly used in IVD research.
- To guide researchers in effectively utilizing IHC for protein expression analysis in IVD tissues.
Main Methods:
- Application of immunohistochemistry (IHC) principles to FFPE IVD tissue.
- Utilizing specific antibody-epitope interactions for antigen localization.
- Development and optimization of IHC protocols for FFPE IVD samples.
Main Results:
- Demonstration of IHC as a viable and common technique for protein detection in IVD tissues.
- Presentation of optimized IHC protocols tailored for FFPE IVD samples.
- Successful localization and semiquantification of protein expression in IVD tissues via IHC.
Conclusions:
- IHC is an essential technique for protein expression analysis in adult human IVD tissues, especially FFPE samples.
- Optimized IHC protocols are critical for reliable antigen detection and research advancement in the IVD field.
- This work provides valuable protocols to facilitate future IVD research utilizing IHC.
Abstract:
Immunohistochemistry (IHC) is a useful technique for the localization and semiquantification of protein expression within tissues. Adult human intervertebral disc (IVD) tissues contain a large amount of auto-fluorescence which often makes immunofluorescence techniques inappropriate on tissue samples but can be applied to isolated cell samples. Thus, IHC remains one of, if not the most common application for protein detection within IVD tissue. Immunostaining localizes antigen expression through specific epitope-antibody interactions. Within the field of IVD research, IHC is commonly used on fresh frozen and paraffin embedded tissues to elucidate the expression of antigens. Here, we discuss the principles of IHC applied to formalin fixed paraffin embedded IVD tissue and supply optimized protocols for antibodies used within our group to guide research within the IVD field.
