Use of Myosin Heavy chain (fast, perinatal) antibody from Sigma-Aldrich on myogenic cell culture

October 03, 2016

Pathology
BWH
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Sigma-Aldrich

Product Name:

Monoclonal Anti-Myosin (Skeletal, Fast) antibody

Catalog Number:

M4276

Browse Similar Products: Myosin Immunofluorescence Antibodies

This antibody was used to analyze the presence of myotubes in differentiated culture of mouse embryonic stem cells (mESCs). This monoclonal anti-Myosin Heavy chain (fast, perinatal) antibody appears highly specific to differentiated muscle lineage, in particular myotubes. Furthermore, staining with this antibody allows to show sarcomeric organization of myofibers as evidenced by the typical striated distribution of the signal at high resolution.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Mouse embryonic stem cells (mESCs), differentiated. Fixed Adherent culture.

Primary Incubation

1:350, overnight, at 4C in Blocking agent, on shaker

Blocking Agent

TBS, 1% FBS, 0.1% TritonX100, 30min, at RT

Secondary Incubation

1:500, AlexaFluor-488-conjugated anti-mouse IgG secondary (Molecular Probes), overnight, at 4C in Blocking agent, on shaker

Tertiary Incubation

N/A

Detection

Fluorescent microscope, FITC/GFP filter for AlexaFluor-488 detection

Results Summary

This antibody works very well on fixed adherent culture containing myogenic cells. High specificity.

Additional Notes

None

Related Categories

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Summary

The Good

Strong specific signal (myotubes and skeletal fibers) by immunofluorescence

The Bad

Signal intensity weaker on human myogenic cultures compared to equivalent mouse cultures

The Bottom Line

Excellent antibody overall. Highly recommended for immunofluorescence study of skeletal myogenesis in mouse. This antibody reacts with Fast Myosin Heavy chain isoforms. Detailed reactivity with developmental MyHCs remains to be analyzed.

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