Cep170 Antibody

Cell Biology
Sloan-Kettering Institute
PHD Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Invitrogen Antibodies

Product Name:

CEP170 Monoclonal Antibody (72-413-1)

Catalog Number:

41-3200

Mature centrioles have assessor structures (called distal and subdistal appendages) attached near their distal ends. My work attempts to elucidate the function of the subdistal appendages of the centriole. Cep170 had been previously reported to localize to both centriole proximal ends and subdistal appendages. We confirmed its localization. Kif2a, another centriolar protein had similar localization. We also needed this reliable antibody to confirm depletion of the proteins with RNAi and CRISPR. The antibodies also allowed us to determine which conditions lead to the mislocalization of these proteins (Cep128, Centriolin, ODF2, Kif2a, cep170, ninein ……).

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

RPE1 cells (human derived). Also works with mouse derived cell lines. Works in 4% PFA fixation of for methanol fixation

Primary Incubation

1:1000 dilution in PBS with 3% BSA and 0.1% triton

Blocking Agent

PBS with 3% BSA and 0.1% triton

Secondary Incubation

Molecular probes anti mouse IgG1 594 secondary antibody in PBS with 3% BSA and 0.1% triton

Tertiary Incubation

DAPI

Detection

Fluorescent microscopy

Results Summary

Cep170 antibody works well for immune fluorescence. Brighter with PFA fixation than with methanol. Brighter signal seen when staining human derived cell lines than those derived from mouse tissue.

DOI or PMID #

27818179

Additional Notes

N/A

Related Categories

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Summary

The Good

It stains the centriole subdistal appendages and centriole proximal ends

The Bad

Quality of stains decline if stored at 4C for over 1 year.

The Bottom Line

This is a reliable Cep170 antibody.

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