Kif2a Antibody Is Sensitive

Cell Biology
Sloan-Kettering Institute
PHD Student

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Abcam

Product Name:

Anti-Kif2a antibody (ab37005)

Catalog Number:

ab37005

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. 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) fixed with either PFA 4% or methanol (-20).

Primary Incubation

1:3000 dilution in PBS, incubate 1 hour with 3% BSA and 0.1% triton

Blocking Agent

PBS with 3% BSA and 0.1 % triton

Secondary Incubation

Anti rabbit Alexa 488 (molecular probes) 1:500, 1 hour with 3% BSA and 0.1% triton

Tertiary Incubation

DAPI

Detection

Immune Fluorescence

Results Summary

When it first arrived, staining was clear at both centrosomes and microtubule cytoskeleton. Centrosome staining remained good regardless of how long the antibody was stored at 4C. Microtubule staining became weaker if antibody was stored at 4C. We aliquoted this antibody at kept at -80. Works in both human and mouse derived cell lines.

DOI or PMID #

27818179

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

It works with nearly any fixation, very sensitive in both human and mouse derived lines

The Bad

Storage

The Bottom Line

It effectively stains Kif2a.

Share your experience with other scientists. Write a Review! »

Join the discussion