Cell Signaling Technology
Smad2 (D43B4) XP® Rabbit mAb
5339
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My goal was to understand interactions between cells and nanoparticles, and the biological effect of those nanoparticles on prostate cancer and non-cancer cells in vitro. To understand if my nanoparticle was triggering the TGF-Beta signaling pathway, I used the Smad2 antibody to stain the protein and check for nuclear translocation.
Immunofluorescence
PNT1A cells (prostate non-cancer) and PC-3 cells (prostate cancer)
Dilution 1:200 in blocking agent, and incubation for 60 minutes at 37 ºC.
Fetal Bovine Serum 10% in PBS
N/A
Widefield fluorescence microscopy (secondary antibody staining with Alexa Fluor 488)
This antibody easily detected Smad2, and I validated that it detects Smad2 translocation to the nuclei by treating the cells with TGF-Beta 3 (10 ng/mL) for 1 hour and proceeding with fixation and staining. It clearly showed Smad2 translocation to the nuclei even in widefield fluorescence microscopy.
https://doi.org/10.1080/17435390.2024.2307616
Images of PC-3 and PNT1A are used to show the differences in staining between cell lines.
It worked every single time, and incubation period was fast.
Too expensive.
Smad2 Antibody has a great signal, even in cells with lower protein expression. I would repurchase it despite the price.