The NRVMs were fixed with 4% paraformaldehyde for 30 min and washed 3 times, each time for 5 min with phosphate buffer saline (1×, PH 7.2–7.4). Next, the cells were permeabilized with 0.2% Triton X-100 for 20 min and blocked with 10% goat serum in PBS for 1h at room temperature. Subsequently, the cells were incubated with mouse α-actinin monoclonal antibody (1:200, Sigma-Aldrich, St Louis, MO, USA) overnight at 4 °C. The cells were washed three times with PBS buffer, then were incubated with FITC labelled secondary IgG antibody (1:200, Jackson, USA) at 37 °C in dark for 2 h. Finally, the cells were stained with DAPI (1:100, Sigma-Aldrich’s Louis, USA) to label the nuclei. To measure cell surface area, the cells were observed by a fluorescence microscope (Carl Zeiss, Oberkochen, Germany). At least 600 cardiomyocytes in 60 fields of view were examined in each group. The cell size was measured by Image J software.
Fitc labelled secondary igg antibody
Manufactured by Jackson ImmunoResearch
Sourced in United States
FITC labelled secondary IgG antibody is a laboratory reagent used in immunoassays and other immunological techniques. It is a secondary antibody that is conjugated with the fluorescent dye FITC (Fluorescein Isothiocyanate). The primary function of this product is to serve as a detection reagent by binding to the constant region of primary antibodies, allowing for visualization and quantification of target analytes.
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2 protocols using fitc labelled secondary igg antibody
1
Immunofluorescence Staining of Cardiomyocytes
2
Immunofluorescence Staining of Neonatal Rat Ventricular Cardiomyocytes
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At the end of cultivation, we discarded the medium and washed NRVMs three times with PBS (1×, pH 7.2–7.4), followed by fixation in 4% paraformaldehyde for 20 min. Cells were then permeabilised with 0.2% Triton X-100 in PBS for 30 min and blocked with 10% goat serum in PBS for 1 h. NRVMs were incubated with a mouse α-actinin monoclonal antibody (1 : 200, Sigma-Aldrich, St. Louis, MO, USA) at 4°C overnight. After three washes with PBS, NRVMs were incubated with an FITC-labelled secondary IgG antibody (1 : 200, Jackson, USA) at 37°C in the dark for 2 h. Finally, 4′,6-diamidino-2-phenylindole (DAPI, 1 : 100, Sigma-Aldrich, St. Louis, USA) was used to label the nuclei. Cell images were captured using a fluorescence microscope (Carl Zeiss, Oberkochen, Germany). For each group, at least 1,000 cardiomyocytes were counted, and the cell surface area was analysed using ImageJ software.
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