NTF97 antibody for Western Blotting

Biochemistry
Stanford University
Senior Research Scientist

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Company:

Abcam

Product Name:

Anti-NTF97/Importin beta antibody

Catalog Number:

ab2811

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, knockdown of hedgehog pathway components was performed and a marker was needed to judge loading of samples for Western Blotting. NTF97 blotting was done to check if this antibody works in 3T3 cells so that it can be used at later stages.

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

Donkey anti-mouse IgG-HRP conjugated; 1:5,000 dilution; 1 hr at room temp.

Tertiary Incubation

None

Detection

ECL regular

Results Summary

NTF-97 was expressed at good levels in 3T3 cells.

Additional Notes

None

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Summary

The Good

Specific signal and no cross-reactive bands were observed

The Bad

None

The Bottom Line

Good NTF97 (Importin-beta) antibody for Western Blotting

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