Common lysosomal markers for immunofluorescence studies are LAMP1, CD63 and lysotracker which can also stain late endosomes. Can anyone suggest a marker/dye that is very specific for lysosomes?
In our hands, Lysotracker was ok but you need live cells. I think you may also try the good "old dye": neutral red.
Good luck
You can try Lysotracker with live cells.
Also, you may want to check a very good publication with a lot of widely accepted advice for studying lysosomal/autophagic function:
https://www.ncbi.nlm.nih.gov/pubmed/26799652
Lysotracker stains acidic organelles and we see that it stains late endosomes as well. My studies require proper distinction between the two which is why Iam looking for something that stains only the lysosomes and not the late endosomes. I came across some stain/dye a while ago which stains lysosomes specifically based on the principle of presence of acid hydrolases present in it but am unable to track the article.
Could you use a CD63 antibody? Although I am not sure if it also labels late endosomes.
A specific marker would be very useful to know about, please keep us posted!
Hi.
CD63 (Lamp3) is more late endosomal and it's true that finding a good specific lysosomal marker is not that easy. A double staining lysotracker + CD63 would allow the discrimination between late endosomes and lysosomes: lysosomes would be lysotracker-positive but almost CD63-negative. Alternatively, you could try to label lysosomal enzymes, such as Cathepsin D, but not sure if there is an good commercial antibody raised against it...
Good luck!
This can be a specific lysosomal marker:
"To functionally assess if LAP increases exposure of L.m. to lysosomal acid hydrolases, we labeled L.m. with 5-dodecanoylaminofluorescein-di-β-D-galactopyranoside (C12FDG) that becomes fluorescent upon cleavage by the lysosomal acid hydrolase β-galactosidase."
http://www.cell.com/cell-host-microbe/abstract/S1931-3128(18)30053-2📷
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