GLI2 Antibody for Immunofluorescence

Biochemistry
Stanford University
Senior Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

R&D Systems

Product Name:

Mouse GLI‑2 Antibody

Catalog Number:

AF3635

I am interested in understanding the mechanism of Hedgehog signal transduction in cultured fibroblast cells. The goal of this experiment is to test if GLI2 protein translocates to the nucleus after treatment of cells with sonic hedgehog ligand. This antibody works for Western blotting but I was curious to test if it works for immunofluorescence.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

NIH3T3 cells

Primary Incubation

1:20 in blocking agent for 1 hr at room temperature

Blocking Agent

1% donkey serum + 1% BSA + 0.1% Triton X-100, in PBS.

Secondary Incubation

Donkey anti-goat Alexa Fluor 488 at 1:1000 in blocking agent for 1 hr at room temp.

Tertiary Incubation

None

Detection

Images were acquired on a Leica TCS SP8 confocal microscope with a 63X oil objective

Results Summary

This antibody detects Gli2 in the cytoplasm at resting conditions. Upon SAG addition for 24 hours (a small molecule that activates hedgehog pathway), overall GLI2 levels increase with a good enrichment in the nucleus.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Detects GLI2

The Bad

Has to be used at very low dilution (1:20)

The Bottom Line

OK for GLI2 staining

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