[IF=11.205] Yingchun Luo. et al. Akkermansia muciniphila prevents cold-related atrial fibrillation in rats by modulation of TMAO induced cardiac pyroptosis. EBIOMEDICINE. 2022 Aug;82:104087  WB ;  Rat.  

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[IF=7.675] Ya-Ling Yin. et al. Citronellal Attenuates Oxidative Stress–Induced Mitochondrial Damage through TRPM2/NHE1 Pathway and Effectively Inhibits Endothelial Dysfunction in Type 2 Diabetes Mellitus. ANTIOXIDANTS-BASEL. 2022 Nov;11(11):2241  IF ;  Rat.  

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[IF=5.923] Chih-Hsin Hsu. et al. miR-29a-3p/THBS2 Axis Regulates PAH-Induced Cardiac Fibrosis. Int J Mol Sci. 2021 Jan;22(19):10574  WB,IHC ;  Human.  

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[IF=2.894] Shen, Zhou. et al. Bone marrow mesenchymal stem cells therapy on bilateral pelvic nerve crush-induced voiding dysfunction in rats. INT UROGYNECOL J. 2022 Apr;:1-8  WB ;  Rat.  

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[IF=2.37] Zhang, Mingming. et al. Both high glucose and phosphate overload promote senescence-associated calcification of vascular muscle cells. INT UROL NEPHROL. Int Urol Nephrol. 2022 Apr;:1-13  IF ;  Mouse.  

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Sample:
Lane 1: A431 cell lysate
Lane 2: Hela cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: mouse heart tissue lysate
Lane 5: mouse smooth muscle tissue lysate
Primary: Anti-alpha smooth muscle Actin (bsm-52396R) at 1:500 dilution
Secondary: Goat Anti-Rabbit IgG - HRP at 1:5000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD

Paraformaldehyde-fixed, paraffin embedded (human tonsil tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-52396R ) at 1:50 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human stomach carcinoma tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-52396R ) at 1:50 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human liver tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-52396R ) at 1:50 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-52396R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-52396R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

RH-35 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-52396R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-52396R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Blank control:Jurkat.
Primary Antibody (green line): Rabbit Anti-alpha smooth muscle Actin antibody (bsm-52396R)
Dilution: 1:50;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1:1000.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.