Although Green Fluorescent, GFP and zsGreen share NO homology, therefore GFP antibodies wont work. As far as antibodies on the market, I have tried clonetech 632474 RAb, and so far it BARELY works in tissue IF. I am currently working on protocols to maintain the fluorescent signal in fixed (4%paraformaldehyde) tissue. I work in the mammary gland, and using a fluorescent, dissection microscope I can see my zsGreen+ structures easily in whole mount.

It is unlikely that you will be able to detect the Zsgreen protein with an antibody to Aequorea-derived GFP. However, there are antibodies available which claim to be used to detect Zsgreen. One such antibdoy is available from Clontech, which they recommend for Western blots. See  link below.

http://www.clontech.com/US/Products/Fluorescent_Proteins_and_Reporters/Fluorescent_Protein_Antibodies/Green_Fluorescent_Proteins

It is difficult to detect GFP in animals models because green light cannot penetrate well into tissue. Please see the article.  http://www.ncbi.nlm.nih.gov/pubmed/17615263

May be you might want to use red-shifted fluroescent protein such as tagrfp. You can also buy it from Everogen.  http://www.evrogen.com/products/TagRFP/TagRFP.shtml

Although Green Fluorescent, GFP and zsGreen share NO homology, therefore GFP antibodies wont work. As far as antibodies on the market, I have tried clonetech 632474 RAb, and so far it BARELY works in tissue IF. I am currently working on protocols to maintain the fluorescent signal in fixed (4%paraformaldehyde) tissue. I work in the mammary gland, and using a fluorescent, dissection microscope I can see my zsGreen+ structures easily in whole mount.

Do you work on fresh whole mount or the ones which are cryo-preserved for endogenous ZS green staining

I have tried it on cryo-preserved and fixed tissues (a variety of methods/antigen retrieval, etc).

Hello everyone,

I 'd like to use an anti-zs green on frozen sections. Is it worth to give a try?

Thanks

 If you are not able to see native zsgreen florescence from your fixed sample, I am afraid that the antibody(632598 Living Colors® ZsGreen Monoclonal Antibody from Clontech) is not going to detect it in western blot. In my experience, the antibody (632598) is a very specific antibody, but the sensitivity is not that great to be comparable to zsgreen1 florescence. 

did you clone anything containing an ATG (preferably with Kozak) in front of the IRES? might be worth trying

I have used anti Zs green which works well

Hi Aparna, can you tell me which (product code) anti zs green works best?

Hi Shiraz Tyebji, I'm using a guinea-pig anti Zsgreen from Frontiers Institute, at 1/1000 (Ref ZsGreen-GP-Af940). It works very well in free-floating, fixed brain sections. They provide this antibody made in rabbit as well if you prefer.

https://w2.frontier-institute.com/w/product-lists/?search=zsgreen

Thanks Vincent Simon i will give that a try.