Kif3a antibody for Western Blotting

Biochemistry
Stanford University
Senior Research Scientist

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Company:

Abcam

Product Name:

Anti-KIF3A antibody

Catalog Number:

ab11259

The hedgehog signaling plays a critical role in embryonic development. It also controls adult tissue homeostasis and deregulation of this pathway results in basal cell carcinoma and medulloblastoma. A unique feature of this pathway is that it requires a tiny antenna like organelle called the primary cilium for signal transduction. My goal is to understand the molecular mechanism of hedgehog signal transduction at the primary cilium. This involves detecting protein-protein interactions of the pathway components, analysis of localization patterns during signal transduction, and transcriptional output measurements. In the present experiment, the ciliary localized kinesin protein, Kif3a, levels were tested by Western Blotting in 3T3 cells with a later goal to knockdown for functional assays.

Experimental Design and Results Summary

Applications

Western Blot

Sample

NIH-3T3 cell lysate

Primary Incubation

1:1000 dilution in blocking buffer; 16 hr at 4 deg

Blocking Agent

5% milk powder in 1xTBS-T(0.1%)

Secondary Incubation

1:10,000 dilution of Donkey anti-rabbit IgG-HRP in blocking buffer; 1 hr at room temp

Tertiary Incubation

None

Detection

ECL pico

Results Summary

Kif3a was expressed in 3T3 cells and the antibody from Abcam picked up a specific band at ~85 kDa marker size.

Additional Notes

None

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Summary

The Good

Specificity of the antibody

The Bad

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The Bottom Line

Recommended for Western Blotting of Kif3a. In addition, this antibody also works for immunofluorescence staining experiments.

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