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Primary rabbit anti mouse α sma antibody

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Primary rabbit anti-mouse α-SMA antibody is a laboratory reagent used to detect the presence of alpha-smooth muscle actin (α-SMA) in mouse samples. It is a polyclonal antibody produced in rabbits and specifically targets the α-SMA protein.

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Lab products found in correlation

Primary rat antimouse f4 80 antibody, by Bio-Rad (1 mentions) Goat anti rabbit igg hrp secondary antibody, by Abcam (1 mentions) Bond polymer refine detection system, by Leica (1 mentions) Vectastain avidin biotin complex conjugated with horseradish peroxidase, by Vector Laboratories (1 mentions)

2 protocols using primary rabbit anti mouse α sma antibody

1

Immunohistochemical Analysis of Macrophages and Myofibroblasts

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Paraffin‐embedded tissue sections were deparaffinized, rehydrated and pretreated with sodium citrate (0.05%) to unmask the antigen, followed by incubation with H2O2 (0.3%) for 25 min at room temperature in dark conditions, to block endogenous peroxidase activity, and with 2% BSA for 20 min at room temperature, to avoid nonspecific binding of the primary antibody. Thereafter, the liver sections were incubated 1 h at room temperature with a primary rat anti‐mouse F4/80 antibody (dilution 1:100; Bio‐Rad, Hercules, CA, USA) followed by incubation for 15 min at room temperature with a biotinylated rabbit anti‐rat IgG secondary antibody (Abcam) and 30 min with HRP goat anti‐rabbit IgG secondary antibody (dilution 1:200; Abcam). For α‐smooth muscle actin (α‐SMA), sections were incubated with primary rabbit anti‐mouse α‐SMA antibody (dilution 1:250; Abcam) for 1 h with HRP goat anti‐rabbit IgG secondary antibody (dilution 1:200; Abcam) for 30 min. The signal was detected by the Bond Polymer Refine Detection system (Leica Biosystems, Wetzlar, Germany). The sections were visualized at 200×, and positive staining for F4/80 and α‐SMA was quantified by histomorphometry. The results are expressed as a percentage of the F4/80‐positive or α‐SMA‐positive area.
Flores‐Costa R., Alcaraz‐Quiles J., Titos E., López‐Vicario C., Casulleras M., Duran‐Güell M., Rius B., Diaz A., Hall K., Shea C., Sarno R., Currie M., Masferrer J.L, & Clària J. (2018). The soluble guanylate cyclase stimulator IW‐1973 prevents inflammation and fibrosis in experimental non‐alcoholic steatohepatitis. British Journal of Pharmacology, 175(6), 953-967.
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2

Immunohistochemical Analysis of Skin Samples

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Multiple 5 μm sections of paraffin-embedded skin samples (mouse or human skin) were cut onto slides, and then de-paraffinized and rehydrated. Immunolabeling of α-smooth muscle actin (α-SMA) and phospho-Smad2 was performed with primary rabbit anti-mouse α-SMA antibody (Abcam) and primary rabbit anti-mouse phospho-Smad2 antibody (Cell Signaling). Immunolabeling of IL-6 was performed with primary rabbit anti-mouse IL-6 antibody or primary rabbit anti-goat IL-6 antibody (Cell Signaling), respectively, using the DakoCytomation system per manufacturer’s instructions. Immunolabeling of CD3 or F4/80 (Abcam) was performed with primary rabbit anti-mouse CD3 or primary rat anti-mouse F4/80, respectively. Appropriate biotinylated secondary antibodies were used, followed by detection with Vectastain avidin/biotin complex conjugated with horseradish peroxidase (Vector Laboratories) and color development with AEC (Dako). Cells positive for α-SMA, pSmad2, CD3 and F4/80 were counted in 10 randomly selected, non-overlapping high-power fields. Blinded sections were photographed with imaging software and IL-6 positive staining was quantified using ImageJ analysis software to calculate the percentage of skin surface area that stained positively for IL-6.
Castelino F.V., Bain G., Pace V.A., Black K.E., George L., Probst C.K., Goulet L., Lafyatis R, & Tager A.M. (2016). An Autotaxin-LPA-IL-6 Amplification Loop Drives Scleroderma Fibrosis. Arthritis & rheumatology (Hoboken, N.J.), 68(12), 2964-2974.
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