Use Of Titin (Ttn, 9D10) Monoclonal Antibody From DSHB on Myogenic Cell Culture

October 31, 2016

Pathology
BWH
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Developmental Studies Hybridoma Bank

Product Name:

9D10 (Titin)

Catalog Number:

9D10-c

 

Browse Similar Products: Titin Immunofluorescence Antibodies

This antibody was used to analyze the presence of muscle fibers in differentiated culture of human pluripotent stem cells (hiPSCs). This monoclonal anti-Titin (Ttnl) antibody appears highly specific to differentiated muscle lineage, in particular myofibers. Furthermore, staining with this antibody allows to show sarcomeric organization of striated muscle.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Human pluripotent stem cells (hiPSCs), 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 skeletal muscle fibers. High specificity.

Additional Notes

Also recognizes cardiomyocytes

Related Categories

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Summary

The Good

Strong specific signal for striated muscle fibers by immunofluorescence

The Bad

Signal intensity somewhat weaker on mouse myogenic cultures compared to human cultures but still acceptable

The Bottom Line

Excellent antibody overall. Highly recommended for immunofluorescence study of myogenesis in human cell cultures. This antibody reacts against Titin which is present in striated muscles- both skeletal muscle and cardiac.

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