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Alexa fluor 594 conjugated donkey polyclonal anti rabbit igg antibody

Manufactured by Jackson ImmunoResearch
Sourced in United States

Alexa Fluor 594-conjugated donkey polyclonal anti-rabbit IgG antibody is a secondary antibody that binds to rabbit IgG antibodies. It is conjugated with the Alexa Fluor 594 fluorescent dye, which enables detection and visualization of target proteins in various applications, such as immunofluorescence microscopy and flow cytometry.

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3 protocols using alexa fluor 594 conjugated donkey polyclonal anti rabbit igg antibody

1

Immunostaining Protocol for Nucleolin

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hCGp7 cells were fixed with 2% paraformaldehyde, permeabilized with 0.2% Triton X-100, and immunostained with 1.0 μg/mL rabbit polyclonal anti-nucleolin antibody (ab22758; Abcam, Cambridge, UK) overnight at 4 °C and then with 0.1 μg/mL Alexa Fluor 594-conjugated donkey polyclonal anti-rabbit IgG antibody (Jackson ImmunoResearch, West Grove, PA, USA) for 1 h at room temperature [6 (link),7 (link),8 (link),10 (link)]. Cell nuclei were stained with DAPI (Nacalai).
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2

Immunocytochemistry Analysis of Myoblasts

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Immunocytochemistry of myoblasts was performed as previously described (Takaya et al., 2017 (link); Nihashi et al., 2019a (link),b (link)). The myoblasts were fixed with 2% paraformaldehyde, permeabilized with 0.2% Triton X-100 (Nacalai), and immunostained with 0.5 μg/ml mouse monoclonal anti-myosin heavy chain (MHC) antibody (MF20; R&D Systems, MN, USA) and 1.0 μg/ml rabbit polyclonal anti-nucleolin antibody (ab22758; Abcam, Cambridge, UK). 0.1 μg/ml each of Alexa Fluor 488-conjugated donkey polyclonal anti-mouse IgG antibody and Alexa Fluor 594-conjugated donkey polyclonal anti-rabbit IgG antibody (Jackson ImmunoResearch, PA, USA) were used as secondary antibodies. Cell nuclei were stained with DAPI (Nacalai). High-resolution fluorescent images were taken under an EVOS FL Auto microscope (AMAFD1000; Thermo Fisher Scientific). The ratio of MHC+ cells was defined as the number of nuclei in the MHC+ cells divided by the total number of nuclei, and the fusion index was defined as the number of nuclei in the multinuclear MHC+ myotubes divided by the total number of nuclei using ImageJ software (National Institutes of Health, USA).
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3

Immunocytochemistry Analysis of Myoblast Differentiation

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Immunocytochemistry of myoblasts was performed as previously described (Takaya et al., 2017; (link)Nihashi et al., 2019a; (link)Nihashi et al., 2019b) .
The myoblasts were fixed with 2% paraformaldehyde, permeabilized with 0.2% Triton X-100, and immunostained with 0.5 μg/ml mouse monoclonal anti-myosin heavy chain (MHC) antibody (MF20; R&D Systems, MN, USA) and 1.0 μg/ml rabbit polyclonal anti-nucleolin antibody (ab22758; Abcam, Cambridge, UK). 0.1 μg/ml each of Alexa Fluor 488-conjugated donkey polyclonal anti-mouse IgG antibody and Alexa Fluor 594-conjugated donkey polyclonal anti-rabbit IgG antibody (Jackson ImmunoResearch, PA, USA) were used as secondary antibodies. Cell nuclei were stained with DAPI (Nacalai). High-resolution fluorescent images were taken under an EVOS FL Auto microscope (AMAFD1000; Thermo Fisher Scientific). The ratio of MHC + cells was defined as the number of nuclei in the MHC + cells divided by the total number of nuclei, and the fusion index was defined as the number of nuclei in the multinuclear MHC + myotubes divided by the total number of nuclei using ImageJ software (National Institutes of Health, USA).
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