Use Of Pax7 Mouse Monoclonal From DSHB On Differentiated Mouse ES Cells

November 01, 2016

Pathology
BWH
Research Scientist

Overall

Quality of Results

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Company:

Developmental Studies Hybridoma Bank

Product Name:

Pax7

Catalog Number:

Pax7-c

This antibody was used to analyze the presence of myogenic progenitors in cultures of differentiated mouse embryonic stem cells (mESCs). This monoclonal anti-Pax7 antibody appears specific to fetal stage myogenic progenitors and mature satellite cells. Some neural rosette-forming progenitors were also positive. Staining with this antibody results in specific nuclear signal as expected for a transcription factor such as paired-box Pax7.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

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

Primary Incubation

1:200, overnight, at 4C in Blocking agent, on shaker

Blocking Agent

Tris-Buffered Saline, 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 well on fixed adherent culture containing fetal myogenic progenitors and also satellite like cells. Signal is nuclear specific.

Additional Notes

Antibody also recognizes some neural rosettes progenitors

Related Categories

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Summary

The Good

Specific staining for fetal myogenic progenitors and satellite, and some neural (rosette) progenitors. Works also on differentiated human PSC cultures.

The Bad

Signal varies from moderate to good intensity

The Bottom Line

Good antibody overall. Specific nuclear staining with moderate to good signal Recommended for immunofluorescence study of differentiating mouse and human pluripotent stem cells cultures into myogenic progenitors.

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