Strong Signal: Anti-Mi2 Immunofluorescence

June 17, 2014

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Biological Science
Florida State University
Assistant Professor

Company:

Santa Cruz Biotechnology, Inc.

Product Name:

Anti-Mi2 (C-17)

Catalog Number:

sc-8774

The purpose of this experiment was to identify an antibody that was suitable for detection of Mi2 in human cells by indirect immunofluorescence.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

human hTERT-BJ1 cells

Primary Incubation

1:200 at room temperature for 60 minutes. Staining was performed in 1 x PBS and 0.1% Tween 20

Blocking Agent

Slide was pre blocked with 3% BSA in 1 x PBS and 0.1% Tween 20

Secondary Incubation

1:100 dilution of FITC Donkey anti-Goat secondary diluted in 1 x PBS and 0.1% Tween-20 for 60 minutes at room temperature

Tertiary Incubation

No tertiary incubation was performed

Detection

Direct labeled Donkey anti-Goat antibodies

Results Summary

The signal was strong and clean and restricted to the nucleus as expected.

Additional Notes

Cells were grown directly on slides and washed briefly with 1 x PBS before fixing and extracting for 10 minutes at room temperature in 4% formaldehyde, 1 x PBS and 0.1% Triton-X100

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Summary

The Good

The signal was strong and as expected for the distribution of this protein

The Bad

No drawbacks were evident with this antibody

The Bottom Line

This antibody is good for immunofluorescence. Santa Cruz provide 1ml of antibody solution which will last a long time.

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