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α sma

Manufactured by Lab Vision
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α-SMA (Alpha-Smooth Muscle Actin) is a protein marker used in immunohistochemistry and Western blotting applications. It is a key component of the contractile apparatus in smooth muscle cells and is often used to identify and characterize these cell types in various tissues.

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3 protocols using α sma

1

Immunohistochemical Tissue Analysis

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Paraffin-embedded sections were rehydrated and antigen retrieval for Ki67 was performed by microwaving in 10 mmol/L sodium citrate (pH 6.0) for 10 minutes. Frozen sections were fixed with acetone and stained for alpha smooth muscle actin (α-SMA) (Sigma-Aldrich, St Louis, MO). α-SMA primary antibodies were incubated at 4 °C overnight, whereas Ki67 (Lab Vision, Freemont, CA) primary antibody was incubated for 1 hour at room temperature. Signal was enhanced using the tyramide amplification system (PerkinElmer, Boston, MA).
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2

Mesenchymal Stem Cell Characterization

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Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin-streptomycin solution, and 0.25% trypsin − 0.02% EDTA solution were purchased from Lonza Company, Swiss. The monoclonal mouse human antibody for CD44, CD 29, CD 34, Alpha smooth muscle actine (α- SMA), Proliferating cell nuclear antigen (PCNA) and biotinylated goat anti-mouse secondary antibody were purchased from LABVISION USA. Carbon tetrachloride (CCL4) (concentration 100%) and olive oil were obtained from Algomhoria Company.
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3

Immunohistochemical Analysis of Tissue Biomarkers

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Tissue sections were deparaffinized and subsequent blockage of the endogenous peroxidase activity was achieved by incubation with 2.5% methanolic hydrogen peroxide for 30 min. The tissue sections were stained with rabbit antibodies against Ki-67, MMP13, collagen I, desmin (GeneTex, Irvine,CA) or TIMP-1 (Proteintech Group Inc, Chicago, USA) by using the Mouse/Rabbit Probe HRP Labeling Kit (BioTnA Biotech, Kaohsiung, Taiwan). Binding of mouse monoclonal antibodies against GNMT (14-1, YMAC Bio Tech, Taiwan), α-SMA (Lab Vision, Fremont, CA) and TGF-β1 (R&D Systems) was detected by using the N-Histofine Mousestain Kit (Nichirei Biosciences, Tokyo, Japan). The immunohistochemical assays were performed according to the manufacturer’s instructions. The sections were developed with 3-3′ diaminobenzidine (DAB) (Dako, Milan, Italy) and finally counterstained with hematoxylin (Dako). Negative controls were performed using normal mouse antiserum instead of the primary antibody.
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