Immunohistochemistry (IHC) were performed by the standard ENVISION technique as described previously [87 (link)]. Briefly, deparaffinised and rehydrated tissue sections were blocked using standard immunoperoxidase procedure before antigen retrieval (30 minutes, 99°C, Tris EDTA buffer with 0.05% Tween-20). Tissues were then incubated with rabbit polyclonal EV-A71 VP1 (GeneTex, USA) at 4°C overnight. After washing, tissues were then incubated with goat-anti rabbit HRP-conjugate (Dako, Denmark) for 30 minutes at room temperature. Tissues were stained using DAB (Dako) and counterstained with hematoxylin (Dako). The tissues were mounted using DPX (Dako) prior to examination under a light microscope. The negative control tissues for IHC included mock-infected ICR mice brains (n = 2) and hind-limb muscle tissues (n = 2). Isotype control antibodies or normal rabbit immunoglobulin fractions (Dako) were also used to exclude non-specific staining.
Dibutylphthalate polystyrene xylene (dpx)
Manufactured by Agilent Technologies
The DPX is a compact and versatile laboratory instrument designed for high-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography (UHPLC) applications. It provides precise and reliable sample injection, column temperature control, and system pressure monitoring to support accurate and reproducible chromatographic analysis.
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2 protocols using dibutylphthalate polystyrene xylene (dpx)
1
Immunohistochemistry Technique for EV-A71 Detection
2
GFAP Immunohistochemistry in mPFC
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Immunohistochemical analysis of GFAP was performed on the sections of the mPFC. Five-µm-thick sections of the mPFC were fixed with 10% phosphate-buffered formalin for 6 hours. After blocking endogenous peroxidase, the sections were incubated with the primary antibodies, polyclonal anti-GFAP (Dako, Glostrup, Denmark) diluted 1:100. The peroxidase reaction was visualized using 0.03% DAB and 0.005% hydrogen peroxide. The immunostained sections were slightly counterstained with Cresyl violet, dehydrated, cleared, and mounted in DPX (Dako).
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