Sections of 4–5 µm thickness were processed for immunohistochemistry as described by [30 (link)]. Briefly, the sections were deparaffinized, rehydrated, and rinsed in tap water. Then the slides were embedded in 3% hydrogen peroxide for 10 minutes, immersed in antigen retrieval solution and later blocking of nonspecific protein binding was done by incubation in 10% normal goat serum in PBS for 1 hours at room temperature. The slides were incubated with the diluted primary antibody against α- smooth muscle actin (Rabbit Polyclonal antibody, Cat: RB-9010-P0; Lab-Vision, Fremont, CA, USA) at dilution of 1:200 for 2 hours to identify mesangial cell proliferation in paraffin sections.
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Immunohistochemical Analysis of Mesangial Cell Proliferation
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Actin
Antibody
Antigen
Cell proliferation
Dilution
Goat
Hydrogen peroxide
Immunohistochemistry
Mesangial
Paraffin
Protein
Rabbit
Serum
Smooth muscle
Vision
Corresponding Organization :
Other organizations : Zagazig University, University of Bisha
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- Immunohistochemistry protocol
- Mesangial cell proliferation
- Sections of 4-5 µm thickness
- Deparaffinization, rehydration, and rinsing in tap water
- Embedding in 3% hydrogen peroxide for 10 minutes
- Antigen retrieval solution
- Blocking of nonspecific protein binding with 10% normal goat serum in PBS for 1 hour at room temperature
- Incubation with primary antibody against α-smooth muscle actin (1:200 dilution) for 2 hours
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