Alpha Tubulin for Immunofluorescence

Food Science and Human Nutrition
Colorado State University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
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Company:

Santa Cruz Biotechnology

Product Name:

Alpha Tubulin

Catalog Number:

sc-5286

This was an experimental project to test whether we could visualize alpha tubulin under our fluorescent microscope.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

H4IIE rat hepatoma cells

Primary Incubation

1:50 antibody, 1% BSA, in PBST (1X PBS+ 0.1% Tween 20) overnight at 4 degrees C.

Blocking Agent

1% BSA, 22.52 mg/ml glycine in PBST (1X PBS+ 0.1% Tween 20) for 30 minutes at room temperature

Secondary Incubation

1:200 Alexa Fluor 555 antibody, 1% BSA, in PBST (1X PBS+ 0.1% Tween 20) for 1 hour at room temperature

Tertiary Incubation

NA

Detection

Olympus BX63, Cy3 fluorescent filter

Results Summary

This antibody was easily detectable with minimal background. I would consider using a higher dilution to save on antibody because this worked so well.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Very bright, very obvious tag on 90-100% of the cells in my sample.

The Bad

None

The Bottom Line

Great antibody for immunofluorescence. Worked the first time with no optimization necessary.

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