IHC was perfomed as described previously with modifications18 (link). FT organoids were released from the Matrigel with cold 4% paraformaldehyde (Fisher Scientific) and fixed for 2 hours before being embedded in Specimen Processing HistoGel™ (Thermo Fisher). The organoids encapsulated in the Histogel were dehydrated in an ascending series of alcohol, followed by isopropanol and acetone at room temperature. The dehydrated organoids were paraffin-embedded and cut into 5 μm sections on a microtome. Hematoxylin & Eosin (H&E) staining and IHC were carried out following standard procedures. The primary antibodies used in IHC included: Mouse anti-E Cadherin (BD, 610181); Rabbit anti-Pax8 (ProteinTech, 10336–1-AP); Mouse anti-acetylated Tubulin (Sigma, T7451).
Specimen processing histogel
Manufactured by Thermo Fisher Scientific
HistoGel™ is a tissue embedding medium designed for histological specimen processing. It is a water-soluble, transparent gel that provides a solid support for tissue samples during sectioning and staining procedures. HistoGel™ solidifies at room temperature, creating a stable matrix to hold the specimen in place.
Automatically generated - may contain errors
2 protocols using specimen processing histogel
1
Immunohistochemical Analysis of Organoids
2
Organoid Immunohistochemistry Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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IHC was perfomed as described previously with modifications 18 . FT organoids were released from the Matrigel with cold 4% paraformaldehyde (Fisher Scientific) and fixed for 2 hours before being embedded in Specimen Processing HistoGel ™ (Thermo Fisher). The organoids encapsulated in the Histogel were dehydrated in an ascending series of alcohol, followed by isopropanol and acetone at room temperature. The dehydrated organoids were paraffin-embedded and cut into 5 μm sections on a microtome. Hematoxylin & Eosin (H&E) staining and IHC were carried out following standard procedures. The primary antibodies used in IHC included: Mouse anti-E Cadherin (BD, 610181); Rabbit anti-Pax8 (ProteinTech, 10336-1-AP); Mouse anti-acetylated Tubulin (Sigma, T7451).
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