BioLegend
Purified anti-HA.11 Epitope Tag Antibody
902301
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I am interested in understanding the mechanism of Hedgehog signaling in cultured fibroblast cells. The goal of this experiment was to test this HA antibody from BioLegend for immunofluorescence staining of an HA-tagged protein that was over-expressed in NIH3T3 cells. The reason was selecting this antibody was that I had used this antibody for staining other HA-tagged proteins in a different cell line.
Immunofluorescence
NIH3T3 cells
1:500 in blocking agent for 1 hr at room temp.
1% donkey serum + 1% BSA + 0.1% Triton X-100, in PBS.
Donkey anti-rabbit 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
IH3T3 cells were seeded on a coverslip and transfected with a HA-tagged Smoothened protein. Confluent cells were serum starved for 24 hours. This step induces primary cilia. Cells were then treated with a Smoothend agonist (SAG) at 100 nM for 4 hr. Primary cilia were stained with an acetylated tubulin along with the HA antibody. HA-tagged Smoothened nicely translocates to primary cilia after SAG treatment.
Clean antibody for immunofluorescence
A wonderful antibody for immunofluorescence staining