Hi, I am working with HEK293 expressed by Dectin-1. I have a problem after fixation with PFA 4%. despite the fact that I am coating the cover glasses with PLL, the cells jump off after fixation. I am wondering if anyone has a recommendation.
Also, I stain the with anti-hDectin1 IgG as a primary and Alexa flour 647 goat-anti mouse as the secondary. I image them in dSTORM. I am looking for Dectin-1 domain but I am not sure what I should see exactly. could you give me an idea about that please.
Hi Akram!
PLL increase cell adhesion a bit different way comparing to natural substrate. Try to cover your coverslips with collagen/gelatin or just FBS. It can help.
If you want to image the cells try using Millicell EZ SLIDES I have a very good expirience with this product the cell adhesion the exact cating is propietary but the cell adhesion and overall expieience with IHC process is very good.
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