Great Antibody Against Pericentrin

Molecular Oncology
Massachusetts General Hospital
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Abcam

Product Name:

Anti-Pericentrin Antibody

Catalog Number:

ab4448

Antibody is really really good and works incredibly well to mark centrosomes. Antibody is on the more expensive side, but antibody works well at 1:2000. This is the protocol used: 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

1:2000 in PBS+1% BSA 1 hour room temp

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

Antibody is very strong and specific. I recommend using Methanol fixation alone or paraformaldehyde + methanol (as was described in the protocol) to keep centrosomes in tact. Overall antibody worked very well for IF

Additional Notes

None

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Summary

The Good

Works great

The Bad

On the expensive side

The Bottom Line

Great centrosome marker

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