Hi folks,
I am recently working with Arabidopsis to look at the protein which has been reported to be at cell plate involved in cytokinesis with YFP tag. However, when I fixed seedlings which functionally expressed this protein fused with myc or YFP tag with 4% paraformaldehyde and used anti-myc antibody (9E10, sigma or C3956, Sigma) or antiYFP antibody (A1112, Invitrogen) to detect signal, I found the proteinwas no longer at cell plate any more, instead, it was only cytosolic (this protein has also shown to be in cytosol). Does anybody have experience with this case? Is there any paper reporting paraformaldehyde can affect localization of proteins? How can I do with immunostaining?
Thank you in advance!