Hi, we want to use an antibody staining to mark the outlines of neurons (these are neurons from adult mice, VNO). I've looked at a few commonly used membrane markers - anti-pan-cadherin antibody (abcam, sigma, CST)and anti-Na-K atpase antibody (abcam, sigma, CST), but haven't found a reference that uses them for IF/IHC specifically in neurons. Can anyone help with this? Has anyone used an antibody to stain neuronal membranes(cultured neurons will also work), and have a reference paper for the same?
Try to get an antibody a/g K channels. Anti-phospholipid antibody is another choice.
Best,\AMB
Hi Abdul, thanks a lot for your help. Are those commonly used in papers? Can you let me know of any reference papers that have used these?
Best
Sourish
Wheat germ agglutinin (WGA) is often used to label plasma membrane. WGA is a lectin that binds selectively to some carbohydrate moiety on glycoproteins, which are abundant on the plasma membrane. We get it from Molecular Probes. Its available in fluorescent form conjugated to a range of AlexaFluor dyes (and probably other dyes as well). We typically use it diluted in the 1:50 range, but you'd probably want to titrate it for your preparation. WGA is NOT neuron-specific, so it will label all the cells in your culture (if you have other kinds of cells in your cultures, you'd need to be able to distinguish them with an additional marker or by morphology). WGA is compatible with immunolabeling procedures. We apply it mixed in with our secondary antibodies. An important point is that lectin binding is Ca++ dependent (antibody binding is not), so whatever buffer formulation you use for your labeling would have to contain a physiologically relevant level of Ca++. We use HBSS for our experiments, but any PBS formulation that contains Ca++ will work.
Good luck.
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