Smoothened Antibody Works

Cell Biology
Sloan-Kettering Institute
PHD Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Abcam

Product Name:

Smoothened Antibody

Catalog Number:

ab38686

This smoothened antibody is pretty good. Signal is not strong and seems to only detect Human smoothened (not mouse, despite company specifications).

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

1:200 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 Rabbit Igg 594 secondary antibody in PBS with 3% BSA and 0.1% triton

Tertiary Incubation

DAPI

Detection

Immunoluorescence

Results Summary

Stained smoothed in RPE-1 cells without SAG induction. Marked cilia with Acetylated tubular in green. Red smoothened can be seen at the tips of some cilia. Antibody works pretty well but has its drawbacks. Signal is not super strong (even with SAG addition). Mouse Smo stains more faintly than human SMO with this antibody.

DOI or PMID #

27818179

Additional Notes

N/A

Related Categories

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Summary

The Good

It works

The Bad

Not the brightest antibody. Not useful for Western Blotting

The Bottom Line

This is a pretty good antibody for immunofluorescence but I could not get it to work with a western blot.

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