Hello,
I am working on the enteric nervous system of adult zebrafish and I am trying to do an EdU-BrdU chase on a GFP transgenic line and the HCl treatment kind of destroys the tissue (gastrointestinal tube). I cannot see the green fluorescence anymore or much fewer cells that normally -even though I stain for GFP again after the treatment- and the tissue looks damaged. Any alternatives?
Thank you,
Elina
Hi Elina,
I am having the same problem with another tissue. Did you maybe figure out a solution? Perhaps longer rebuffering?
Best,
Tijana
I am wondering if you two have solved the issue or not?
Actually I have the same issue with you two. I want to do co-staining of BrdU with DCX,GFAP and NeuN.
Louis
Hi both... unfortunately I haven't solved the issue as yet. I will be trying another BrdU antibody that doesn't require HCl but I am currently on leave and didn't have the chance yet.
Dear Evangelia,
Could you share the reference for that antibody? Many thanks for sharing information here.
Yes! You can try this one: DHSB G3G4! Let me know how that worked 😀 I havent tried it yet.
And how did you do antigen retrieval if not with HCl? :) On the official page of the antibody they still state use HCl. Was it citrate buffer?
Hi! I am having a similar issue with BrdU stainings on mouse intestine, but I have just found this paper:
Article Antigen-Retrieval Procedure for Bromodeoxyuridine Immunolabe...
As a very brief summary, they claim that a proper antigen retrieval step in citrate buffer with a stable temperature is able to substitute for the need to treat the samples with HCl. I still have to give it a try, but it might be worth a shot.
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