Immunostaining of Skeletal Tissues
Immunohistochemistry (IHC) is a routinely used technique in clinical diagnosis of pathological conditions and in basic and translational research. It combines anatomical, immunological, and biochemical methods and relies on the specific binding of an antibody to an antigen. Using the technique with mineralized tissues is more challenging than with soft tissues. Demineralizing the samples allows for embedding in paraffin wax, and also facilitates cryosectioning. This chapter describes methods for IHC on formaldehyde-fixed, demineralized, paraffin-embedded, or frozen sections to detect antigens in skeletal tissues.
Key wordsImmunohistochemistry Immunofluorescence Paraffin section Cryosection Antigen retrieval Skeletal tissues Demineralization.
This chapter is dedicated to the memory of Dr. Tobias Kurth, who wrote the previous edition. We are grateful for his invaluable contribution to optimizing the protocols described herein. We thank the staff of the Microscopy and Histology Facility at the Institute of Medical Sciences for their support, and Susan Clark for critical reviewing of the manuscript. We are grateful for support to our research from Arthritis Research UK (grants 19271, 19429, 20050, 20775), the Medical Research Council (grants MR/L020211/1, MR/L022893/1), and Tenovus Scotland (grant G13/14).
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