Acetylated Tubulin Antibody for Immunofluorescence Staining

Biochemistry
Stanford University
Senior Research Scientist

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Company:

Sigma-Aldrich

Product Name:

Monoclonal Anti-Tubulin, acetylated antibody produced in mouse

Catalog Number:

T6793

Primary cilia are microtubule based structures that range from 1-15 microns and are present in almost every cell in our body. They have signaling hubs for various pathways and most importantly the sonic hedgehog pathway. I am interested in understanding the mechanism of hedgehog signal transduction. I routinely perform immunofluorescence staining to study trafficking of hedgehog signaling components in response to pathway stimulation. To visualize primary cilia, acetylated tubulin antibody is widely used and I had selected this marker based on previous publications.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

NIH3T3 cells

Primary Incubation

1:3000 dilution in blocking agent for 1 hr at room temp.

Blocking Agent

Blocking buffer: 1% donkey serum + 1% BSA + 0.1% Triton X-100, in PBS.

Secondary Incubation

Donkey anti-mouse Alexa Fluor 647 at 1:1000 dilution in blocking agent for 1 hr at room temp.

Tertiary Incubation

NA

Detection

Images were acquired on a Leica TCS SP8 confocal microscope with a 63X oil immersion objective.

Results Summary

Confluent NIH3T3 cells were serum starved for 24 hours to induce primary cilia. Cells were fixed in 4% PFA. This antibody nicely stains primary cilia.

Additional Notes

None

Related Categories

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Summary

The Good

A nice marker for visualization of primary cilia.

The Bad

None

The Bottom Line

Very good marker for primary cilia

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