Hello everyone,
I would like to get some recommendations for a good antibody that is not phosphorylated Tau for axonal staining only, not for dendrites in primary hippocampal neuronal culture.
Thanks a lot.
Tau is majorly presented in axons, so you dont have worry about dendrities, and antibodies for native tau, I would recommend, Santa cruz, because its reasonably good in all aspects. http://www.scbt.com/datasheet-390476-tau-a-10-antibody.html
Hey, Yogesh. Thanks for the answer however I think, there have been issue that in situ, anti-tau-1 has a stringent specificity for the axons of neurons. The antibody does not stain the cell bodies or dendrites of neurons, nor does it stain any other cell type. However, this in vivo intracellular specificity is not maintained in culture: anti-tau-1 stains the axon, cell bodies, and dendrites of rat hippocampal neurons grown in culture . The specificity of anti-tau-1 was originally thought to represent the restricted expression of tau to axons. Later studies revealed that this specificity is dependant on the state of phosphorylation. In dephosphorylated samples (samples treated with alkaline phosphatase) anti-tau-1 stains astrocytes, perineuronal glial cells, and the axons, cell bodies and dendrites of neurons, while in untreated samples, anti-tau-1 stains only axons. That is the reason why I was specifically looking for non-phosphorylated tau antibody.
As you describe it, Abhiyan, it sounds like the anti-tau-1 antibody was indeed acting as a non-phospho-specific antibody. In general, companies do not intentionally sell non-phospho-specific antibodies. Some companies sell phospho-antibodies and at the same time have made the corresponding total antibody to the same epitope. Many of these will indeed be non-phospho-specific to some degree.
At the link below, for example, you can see many antibodies for:
Anti Tau (Ab-181)
Anti Tau (Ab-205)
etc.
which are the result of immunizing with the non-phosphorylated peptide.
http://www.tebu-bio.com/search/?contexte=&autocomplete=0&type_contexte=0&search=anti+tau+antibody
Thanks Mark for your suggestions. I would try to check those suggested antibodies. Greetings form Muenster.
Under normal, adult brain conditions, tau is mostly the axons, not in the dendrites.The problem with neuronal cultures of E18 rats or mice, is that tau is everywhere. It takes a long time in culture to eventually have an adult phenotype for tau localization. The workaround, as mentioned above, is to use the Tau-1 mAb. The Tau-1 antibody was first described by Binder et al. in 1985 (JCB), it recognizes tau dephosphorylated at S195, S198, S199, and S202 (Szendrei et al., 1993). Depending on the stage of your cultured neurons, Tau-1 signal will be stronger in the axons, and especially in the distal part of the axon. Please read the work of Mandell & Banker (J. Neuroscience, 1996), to have a better picture of tau localization with this antibody.
Hello Everyone, it's a bit of an old thread but Prof Emmanuel Planel apart from clone PC1C6 could you recommend a good reliable antibody from a host other than mouse (preferably rabbit)? I just need an axonal marker for tissue (rat)? Many thanks!
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