Abcam
Anti-Kif2a antibody (ab37005)
ab37005
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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 ……).
Immunofluorescence
RPE1 cells (human derived) fixed with either PFA 4% or methanol (-20).
1:3000 dilution in PBS, incubate 1 hour with 3% BSA and 0.1% triton
PBS with 3% BSA and 0.1 % triton
Anti rabbit Alexa 488 (molecular probes) 1:500, 1 hour with 3% BSA and 0.1% triton
DAPI
Immune Fluorescence
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.
27818179
N/A
It works with nearly any fixation, very sensitive in both human and mouse derived lines
Storage
It effectively stains Kif2a.