Good Antibody Against Survivin

Molecular Oncology
Massachusetts General Hospital
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Santa Cruz Biotechnology

Product Name:

Survivin Antibody (D-8)

Catalog Number:

sc-17779

Survivin is part of the CPC, and this antibody was used in IF experiments to look at components of the CPC in mitosis. Overall the antibody is easy to use and works fairly well. Best if used overnight at 4C. Protocol is below: Cells were fixed in 3.5% paraformaldehyde for 15 minutes at room temperature. 3 minutes in ice-cold methanol. 5 minutes in PBS+0.5% Triton X5 minutes in PBS 10 minutes in PBS+3% BSA Antibody was incubated in PBS+1% BSA and coverslips incubated overnight at 4C2 - 5 minute washes in PBS Secondary antibodies from Jackson Laboratories (anti-rabbit) was incubated in PBS+1% BSA and coverslips 2 - 5 minute washes in PBS Mounted coverslips using ProLong Gold

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

RPE-1 cells

Primary Incubation

1:1000 in PBS+1% BSA overnight at 4C

Blocking Agent

PBS+3%BSA

Secondary Incubation

1:1000 in PBS+1%BSA for 1 hour

Tertiary Incubation

None

Detection

Florescence imaging was done on a 60X objective

Results Summary

Immunofluorescence images were collected using a 60X objective on a Nikon microscope. Cells were co-stained with DAPI (blue).

Additional Notes

None

Related Categories

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Summary

The Good

It works well

The Bad

For best signal should be incubated at 4C overnight

The Bottom Line

Good antibody to look at Survivin. Seems to be specific on Westerns as well (not shown here).

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