Has anyone blotted for NeonGreen on a western blot and found an antibody that works well?
I haven't tried it yet but I noticed this one from Chromotek recently came on the market: http://www.chromotek.com/products/conventional-antibodies/mneongreen-antibody-32f6/
In my experiments, the one from Chromotek is good to detect mNeon green from overexpressing system. However, it is not sensitive enough to detect fusion proteins at endogenous level. You also need to use very high concentration of Ab (>1/500) and incubate for longer time for WT analysis.
I've used the mouse anti mNG from Chromoteck several times in western blotts within the past two years against whole protein extracts from Arabidopsis seedling tissues. It works really well, high signal and high specificity, in my experience.
Also I would NOT recommend the rabbit anti mNG antibody from NEB/cell technologies. Its terrible and doesn't detect well.
Jan Xue thank you for the answer! What is a dilution you are using for the antibody in WB? Did you use ChromoTek mNeonGreen Monoclonal antibody [32F6]?
https://www.ptglab.com/products/mNeonGreen-antibody-32F6.htm
Tjasa Kosir I was looking at an overexpressed mNG tagged protein in plant tissue. For 0.01g of sample in 10% gel wet-blotted onto nitrocellulose 0.45um, incubated with 3/5000 chromotek mouse anti-mNG overnight 4C and next day goat anti mouse conjugated with HRP 1/4000 dilution. got very strong band where I expected it over several samples and replicates.
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