Recommended cilia marker for immunofluorencence staining

February 17, 2015

Cancer Center
Children's National Health System
Lab Manager

Overall

Quality of Results

Ease-of-Optimization

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Company:

Sigma-Aldrich

Product Name:

Monoclonal Anti-α-Tubulin antibody produced in mouse

Catalog Number:

T5168

The aim of this study was to examine the cilia protein using a fluorencence microcope. I chose this antibody as a cilia marker because it alway gives me great quality images and consistent results.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Mouse inner medullary collecting duct (IMCD) cell line

Primary Incubation

1:2000 diluted in blocking buffer for 1 hour at room temperature

Blocking Agent

1% BSA in 1 x PBST (0.01% tween-20)

Secondary Incubation

Alexa Fluor 594 Donkey Anti-Mouse 1:5000 dulited in blocking buffer, incubate for 1 hour at room temperature

Tertiary Incubation

NA

Detection

Sample mounted in ProLong Gold Antifade Mountant with DAPI (Life Techonoliges #P36941 ) and sample was analyzed on a Olympus FV1000 laser scanning confocal microscope.

Results Summary

Image is taken with a 60X oil lens and analyzed with Olympus Fluoview 3.0 softward. As expected, monoclonal anti-α-tubulin antibody is only stained in IMCD cell cilium (showing red in the image). Scale bar = 50 micrometer.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Gives less backgound and consistent results

The Bad

Price is a bit high compared to similar products

The Bottom Line

This product is the best antibody to mark cilia that I have used. I would recommend this for people who work in the cilia image field.

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