Use Of MyoD (5.8A) Monoclonal Antibody From Santa Cruz Biotechnology On Myogenic Cell Culture

November 02, 2016

Pathology
BWH
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Santa Cruz Biotechnology

Product Name:

MyoD Antibody (5.8A)

Catalog Number:

sc-32758

This antibody was used to analyze the presence of myoblasts in differentiated culture of mouse embryonic stem cells (mESCs). This monoclonal anti-MyoD (5.8A) antibody appears highly specific to differentiated muscle lineage including myoblasts and fibers myonuclei. The staining is specific to the nuclei as expected for a bHlH transcription factor.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

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

Primary Incubation

1:250, overnight, at 4C in Blocking agent, on shaker

Blocking Agent

TBS, 1% FBS, 0.1% TritonX100, 30min, at Room temperature

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 skeletal myoblasts. It also reacts with myonuclei within myotubes. High specificity but signal sometimes weak.

Additional Notes

None

Related Categories

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Summary

The Good

Specific signal (myoblasts and myonuclei within fibers) by immunofluorescence

The Bad

Signal can be moderately weak

The Bottom Line

Good antibody overall. Recommended for immunofluorescence study of skeletal myogenesis in vitro. Works on both mouse and human PSCs-derived myogenic cultures.

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