Anti α sma epr5368

Manufactured by Abcam

Anti-α-SMA EPR5368 is a monoclonal antibody that recognizes the alpha-smooth muscle actin (α-SMA) protein. α-SMA is a cytoskeletal protein commonly used as a marker for smooth muscle cells and myofibroblasts. This antibody can be used to detect and localize α-SMA in various biological samples.

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2 protocols using anti α sma epr5368

Protein Expression Analysis in Liver Tissue

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Liver tissues and cells lysates were prepared with RIPA buffer containing 1× EDTA/proteinase-phosphatase inhibitor cocktail (Pierce). The lysate supernatant was stored at −80 °C until used for immunoblotting. Protein extracts were separated by SDS-PAGE electrophoresis and blotted onto nitrocellulose membrane. Blots were incubated overnight with the following primary detection antibodies: anti-α-SMA EPR5368, (1:2000, Abcam); anti-GAPDH GA1R and anti-β-actin (1:10000 ThermoFisher Scientific). The blots were then incubated with IRDye^® IgG secondary antibody conjugates (Li-Cor) and then revealed using an Odyssey^® gel documentation system following the manufacturer’s instruction (Li-Cor).

Mimche P.N., Lee C.M., Mimche S.M., Thapa M., Grakoui A., Henkemeyer M, & Lamb T.J. (2018). EphB2 receptor tyrosine kinase promotes hepatic fibrogenesis in mice via activation of hepatic stellate cells. Scientific Reports, 8, 2532.

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Multiparametric Flow Cytometry Analysis

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Single cell suspension was obtained as previously described (34 (link)). The cells were stained with antibodies, purchased from either BioLegend [anti-CD3e (145-2C11, RRID: AB_312660), anti-CD8𝛼 (53-6.7, RRID: AB_2888883), anti-CD11b (M1/70, RRID: AB_312791), anti-CD25 (PC61.5, RRID: AB_312847), anti-CD29 (HMβ1-1, RRID: AB_312885), anti-CD31 (390, RRID: AB_10613644), anti-CD45 (30-F11, RRID: AB_2563598), anti-CD140a (PDGFRα, RRID: AB_1953268, APA5), anti-CD324 (EpCAM, G8.8, RRID: AB_1134172), anti-VEGFR3 (AFL4, AB_10680790), and anti-PDPN (8.1.1, RRID: AB_2629802)] or from Thermo Fisher Scientific [anti-CD4 (RM4-5, RRID: AB_464902), anti-CD69 (H1.2F3, RRID: AB_1210795), anti-FoxP3 (FJK-16s, RRID:AB_467575), and anti-NK1.1 (PK136, RRID: AB_2536075)] or from Abcam [anti-αSMA (EPR5368, RRID: AB_11129103)]. Cells were also stained with Fixable Viability Dye (Thermo Fisher Scientific) and dead cells were gated out from the analysis. Cell types were determined as following; lymphatic cells: CD45⁺/CD3⁺, CD8 T⁺ cells: CD45⁺/CD3⁺/CD8⁺, Tregs: CD45⁺/CD3⁺/CD4⁺/FoxP3⁺, cancer cells (MOC1): CD45⁻/ CD31⁻/EpCAM⁺/CD140a⁻/VEGFR3⁻, cancer cells (MC38-luc): CD45⁻/CD31⁻/CD29⁺/αSMA⁻, CAFs (MOC1): CD45⁻/CD31⁻/EpCAM⁻/CD140a⁺/VEGFR3⁻, CAFs (MC38-luc): CD45⁻/CD31⁻/αSMA⁺, vascular endothelial cells: CD45⁻/CD31⁺/EpCAM⁻/CD140a⁻/VEGFR3⁻, lymphatic endothelial cells: CD45⁻/CD31⁻/EpCAM⁻/CD140a⁻/VEGFR3⁺.

Kato T., Furusawa A., Okada R., Inagaki F., Wakiyama H., Furumoto H., Fukushima H., Okuyama S., Choyke P.L, & Kobayashi H. (2023). Near-Infrared Photoimmunotherapy Targeting Podoplanin-Expressing Cancer Cells and Cancer-Associated Fibroblasts. Molecular cancer therapeutics, 22(1), 75-88.

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