Purification of Anti-OSH1 IgG and Immunohistochemical Staining of Rice Shoot Apical Meristem

Methods Mol Biol. 2025:2869:61-73. doi: 10.1007/978-1-0716-4204-7_8.

Abstract

Immunohistochemical staining (IHC) enables the visualization of protein localization within tissues and organs. Here, we describe IHC to detect the ORYZA SATIVA HOMEOBOX1 (OSH1) protein in sections of rice shoot apical meristems (SAMs). First, anti-OSH1 IgG is purified from OSH1 polyclonal rabbit antisera. For subsequent IHC, rice shoot apices are fixed and embedded in paraffin and sections are prepared for IHC. In this IHC procedure, two different labeling methods, fluorescein isothiocyanate and alkaline phosphatase, are utilized for secondary antibody detection.

Keywords: Alkaline phosphatase; FITC; IHC; IgG purification; Paraffin section.

MeSH terms

  • Animals
  • Homeodomain Proteins / metabolism
  • Immunoglobulin G* / isolation & purification
  • Immunoglobulin G* / metabolism
  • Immunohistochemistry* / methods
  • Meristem* / metabolism
  • Oryza* / metabolism
  • Plant Proteins* / isolation & purification
  • Plant Proteins* / metabolism
  • Plant Shoots / metabolism
  • Rabbits
  • Staining and Labeling / methods

Substances

  • Immunoglobulin G
  • Plant Proteins
  • Homeodomain Proteins