After fixing with 4% formaldehyde (Sigma-Aldrich; Merck KGaA) at 4˚C for 30 min, the cells were permeabilized with 0.1% Triton X-100 in PBS and then blocked with 10% normal goat serum (Invitrogen; Thermo Fisher Scientific, Inc.) for 1 h at room temperature. The cells were subsequently incubated with primary antibodies against NOX4 (1:100; cat. no. PA5-72816; Thermo Fisher Scientific, Inc.) at 4˚C overnight, followed by FITC-conjugated anti-rabbit IgG secondary antibody (1:100) for 1 h at room temperature (cat. no. A30008; Invitrogen; Thermo Fisher Scientific, Inc.). Fluorescence was assessed by fluorescence microscopy (magnification, x400; Leica Microsystems GmbH). Quantification of fluorescence intensity or positive cell/nuclear ratios was performed using ImageJ version 1.47 (National Institutes of Health). The results are presented as the mean ± standard error of the mean (SEM) from ≥3 independent experiments.
Fitc conjugated anti rabbit igg secondary antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The FITC-conjugated anti-rabbit IgG secondary antibody is a laboratory reagent used in various immunoassay techniques. It is designed to detect and bind to rabbit primary antibodies, with the fluorescent FITC label allowing for visualization and detection.
Automatically generated - may contain errors
2 protocols using fitc conjugated anti rabbit igg secondary antibody
1
Immunofluorescence Assay for NOX4 Detection
2
Immunohistochemical Analysis of Galectin-1 in Trichinella spiralis Muscle Larvae
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Live T. spiralis muscle larvae were fixed in cold ethanol and xylene and embedded in paraffin for histological analysis. Five-micron sections were deparaffinized in xylene, rehydrated with a graded alcohol series, washed and incubated with rabbit anti-human galectin-1 affinity-purified polyclonal antibody (1:1000) (Institute for the Application of Nuclear Energy - INEP, Belgrade, Serbia), overnight at 4 °C. After washing, sections were incubated with FITC-conjugated anti-rabbit IgG secondary antibody (1:1500) (Molecular probes, Thermo Fisher scientific, Waltham, MA, USA) or with biotin-labeled goat anti-rabbit IgG secondary antibody (1:500) (Vector Laboratories, Burlingame, CA, USA), for 1 h at room temperature (RT). Non-specific binding was assessed by omitting the anti-human galectin-1 incubation step. Indirect immunofluorescence (IIF) staining was examined by ultraviolet microscopy (AXIO Imager A1, Carl Zeiss AG, Goettingen, Germany). Incubation with biotin-labeled goat anti-rabbit IgG was followed by ABC (avidin/biotinylated horseradish peroxidase complex, Vector) (1:1000) for 30 min, and the reaction was developed by adding the chromogenic substrate. The stained sections were viewed using light microscopy (AXIO Imager A1, Carl Zeiss AG).
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