The immunofluorescence staining of GPR30 and p-eNOS in the HUVECs was performed as previously described (23 (link)). Following treatment, the HUVECs were fixed in 4% formaldehyde (Aladdin, Shanghai, China) and blocked with 10% normal goat serum (Sigma-Aldrich), then incubated with anti-GPR30 antibody (1:80 dilution) or anti-p-eNOS Ser1177 antibody (1:25 dilution). A fluorescein isothiocyanate-conjugated goat anti-rabbit antibody (1:50 dilution; sc-2012; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) was then used. The nuclei were stained with propidium iodide (3 mg/ml; Santa Cruz Biotechnology, Inc.). Images were acquired using a confocal microscope (FV10i; Olympus Corp., Tokyo, Japan).
Fluorescein isothiocyanate conjugated goat anti rabbit antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Fluorescein isothiocyanate-conjugated goat anti-rabbit antibody is a secondary antibody conjugated with the fluorescent dye fluorescein isothiocyanate. It is designed to specifically bind to and detect rabbit primary antibodies in various immunoassay techniques.
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3 protocols using fluorescein isothiocyanate conjugated goat anti rabbit antibody
1
Immunofluorescence Staining of GPR30 and p-eNOS in HUVECs
2
Immunostaining of GPR30 and p-eNOS in HUVECs
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The immunofluorescence staining of GPR30 and p-eNOS in HUVECs was performed as previously described [27] . After treatment, cells were fixed in 4% formaldehyde (Aladdin) and blocked with 10% normal goat serum (Sigma), then incubated with anti-GPR30 antibody (1:80 dilution, #ab154069, Abcam) or anti-p-eNOS Ser 1177 antibody (1:25 dilution; #5880; Cell Signaling Technology). A fluorescein isothiocyanate-conjugated goat anti-rabbit antibody (1:50 dilution; Santa Cruz Biotechnology) was then applied. Propidium iodide (3 mg/mL; Santa-cruz) was used to visualise the nucleus. Images were acquired by confocal microscopy (FV10i; Olympus; Japan).
3
GPR30 Immunofluorescence Staining in HTR8/SVneo
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The immunofluorescence staining of GPR30 in HTR8/SVneo was performed as previously described [24] . After treatment, the cells were fixed with formaldehyde (4%) and blocked with 10% normal goat serum (Sigma, St. Louis, MO, USA), then incubated with anti-GPR30 antibody for 48 hours (1:250, Abcam, ab154069). A fluorescein isothiocyanate-conjugated goat anti-rabbit antibody (1:50 Santa Cruz Biotechnology, CA, USA) was then applied for 1 hour at room temperature.
Presidium iodide (3mg/mL) was used as a nuclear stain. Images were acquired by confocal microscopy (FV10i; Olympus, Japan).
Presidium iodide (3mg/mL) was used as a nuclear stain. Images were acquired by confocal microscopy (FV10i; Olympus, Japan).
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