Pax3 Mouse Monoclonal From DSHB On Differentiated Mouse ES Cells

October 12, 2016

Pathology
BWH
Research Scientist

Overall

Quality of Results

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Company:

Developmental Studies Hybridoma Bank

Product Name:

Pax3

Catalog Number:

Pax3-c

Browse Similar Products: Pax3 Immunofluorescence Antibodies

This antibody was used to analyze the presence of neuronal and mesodermal progenitors differentiated from mouse embryonic stem cells (mESCs). This monoclonal anti-Pax3 antibody appears specific to neural rosette-forming progenitors as well as somitic and dermomyotome progenitors.Staining with this antibody results in specific nuclear signal as expected for a transcription factor such as paired-box Pax3.

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

Tris-Buffered Saline, 1% FBS, 0.1% TritonX100, 30min, at room temperature

Secondary Incubation

1:500, AlexaFluor-594-conjugated anti-mouse IgG secondary (Molecular Probes), overnight, at 4C in Blocking agent, on shaker

Tertiary Incubation

N/A

Detection

Fluorescent microscope, TexasRed filter for AlexaFluor-594 detection

Results Summary

This antibody works well on fixed adherent culture containing neural rosettes and somitic progenitors. Neural cells give stronger signal with this antibody.Signal is nuclear specific.

Additional Notes

None

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Summary

The Good

Specific staining for some neural (rosette) and mesodermal 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.

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