R&D Systems
Mouse GLI‑2 Antibody
AF3635
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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.
Immunofluorescence
NIH3T3 cells
1:20 in blocking agent for 1 hr at room temperature
1% donkey serum + 1% BSA + 0.1% Triton X-100, in PBS.
Donkey anti-goat Alexa Fluor 488 at 1:1000 in blocking agent for 1 hr at room temp.
None
Images were acquired on a Leica TCS SP8 confocal microscope with a 63X oil objective
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.
Detects GLI2
Has to be used at very low dilution (1:20)
OK for GLI2 staining