HEK293 cells, normal and immortalized human lung pericytes grown on 12-well plates coated with 0.2% gelatin were fixed with 4% PFA for 15 min at room temperature and then washed with DPBS. After blocking with 1% BSA for 15 min at room temperature, cells were incubated overnight at 4°C with a rabbit monoclonal PDGFR-β antibody (clone Y92; Abcam, Cambridge MA) or a rabbit polyclonal NG2 antibody (H-300; Santa Cruz Biotechnology, Dallas, Texas). Cells were washed with DPBS and incubated for 1h at room temperature with Alexa Fluor 594 goat anti-rabbit IgG (H+L) secondary antibody (Invitrogen, Carlsbad, CA), diluted 1:200 in DPBS. After washing with DPBS, images were acquired using a Zeiss Observer.Z1 microscope with Zen 2 software.
Alexa fluor 594 goat anti rabbit igg h l secondary antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 594 goat anti-rabbit IgG (H+L) secondary antibody is a fluorescently labeled antibody that binds to the constant regions (heavy and light chains) of rabbit immunoglobulin G (IgG) molecules. This secondary antibody is designed for use in various immunodetection techniques, such as immunofluorescence microscopy, flow cytometry, and Western blotting.
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3 protocols using alexa fluor 594 goat anti rabbit igg h l secondary antibody
1
Immunofluorescence Characterization of Pericytes
2
Immunofluorescence Characterization of Pericytes
3
NF-κB p65 Translocation Assay
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LO2 cells grown on glass coverslips were stimulated with 5 μg/ml CCN1 for 60 min. After 4% paraformaldehyde-fixed, 0.5% Triton-X-100-permeablized and PBS/5% BSA-PBS blocked, the cells were stained overnight with anti-NF-κB p65 antibody (Cell Signaling Technology, Danvers, MA, USA), and incubated for 1 additional hour with an Alexa Fluor 594 goat anti-rabbit IgG (H +L) secondary antibody (Invitrogen, Carlsbad, CA, USA). After washing, nuclei were counterstained with DAPI for 20 min. The results were examined using fluorescence confocal microscope. Image acquisition and processing were performed using Zen software (Carl Zeiss Microscopy GmbH, Jena, Germany).
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