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).
Alexa fluor 488 conjugated donkey polyclonal anti mouse igg antibody
Manufactured by Jackson ImmunoResearch
Sourced in United States
Alexa Fluor 488-conjugated donkey polyclonal anti-mouse IgG antibody is a secondary antibody that binds to mouse immunoglobulin G (IgG). The antibody is conjugated to the Alexa Fluor 488 fluorescent dye, which can be detected using appropriate fluorescence detection methods.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Nacalai Tesque (3 mentions)
Evos fl auto microscope, by Thermo Fisher Scientific (3 mentions)
Triton x 100, by Nacalai Tesque (2 mentions)
Ab22758, by Abcam (2 mentions)
Alexa fluor 594 conjugated donkey polyclonal anti rabbit igg antibody, by Jackson ImmunoResearch (2 mentions)
3 protocols using alexa fluor 488 conjugated donkey polyclonal anti mouse igg antibody
1
Immunocytochemistry Analysis of Myoblasts
2
Immunostaining and Analysis of Myoblast Fusion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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). 0.1 μg/ml of Alexa Fluor 488-conjugated donkey polyclonal anti-mouse IgG antibody (Jackson ImmunoResearch, PA, USA) was used as the secondary antibody. Cell nuclei were stained with DAPI (Nacalai).
Fluorescent images were captured using EVOS FL Auto microscope (AMAFD1000; Thermo Fisher Scientific, MA, USA). The ratio of MHC + cells was defined as the number of nuclei in all MHC + cells divided by the total number of nuclei, and the fusion index was defined as the number of nuclei in multinuclear MHC + myotubes divided by the total number of nuclei, which were calculated using the ImageJ software (National Institutes of Health, USA).
Fluorescent images were captured using EVOS FL Auto microscope (AMAFD1000; Thermo Fisher Scientific, MA, USA). The ratio of MHC + cells was defined as the number of nuclei in all MHC + cells divided by the total number of nuclei, and the fusion index was defined as the number of nuclei in multinuclear MHC + myotubes divided by the total number of nuclei, which were calculated using the ImageJ software (National Institutes of Health, USA).
3
Immunocytochemistry Analysis of Myoblast Differentiation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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).
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).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!