Immunofluorescent Analysis of Histone H3 Citrullination
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Other organizations : Nanjing Medical University, Jiangsu Province Hospital
Variable analysis
- Tissue samples were immobilized with OCT at -80°C for ≥24 h
- Expression of Histone H3 (citrulline R2+R8+R17)
- Tissue sections were fixed in 4°C acetone and rehydrated in PBS
- Slides were gently washed in PBS for three times
- Tissue sections were blocked with 5% goat serum at 37°C to reduce non-specific binding
- Tissue sections were stained with Histone H3 (citrulline R2+R8+R17) antibody (1:300 diluted) overnight at 4°C in the dark
- Tissue sections were stained with the fluorescent-conjugated secondary antibody [Goat anti-Rabbit IgG (H+L) Alexa Fluor® 555 conjugate] at 37°C for 60 min in the dark
- Tissue sections were incubated with SYTOX Green diluted in PBS (1:2,000) at 37°C for 15 min
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