Anti-MAP2 antibody - Neuronal Marker (ab5392)

Dear Mr. or Ms.,

One of my costumers wants to know:

1-) Can antibody Anti-MAP2 (ABCAM - Code ab5392) be used as a marker for myenteric neurons in rat and mouse specimen?

2-) If that's possible, which one do you consider the best choice for marking myenteric neurons (total population): anti-MAP2 (ab5392) or anti-PGP9.5 (ab8189)?

I'm forward to hearing from you.

Thanks for your attention.


Best Regards,

ANSWER:

Thank you for your enquiry.

1. I have done a short literature search and can confirm that the MAP2 protein is a good marker for myeneric neurons as well as neurons in general. However, I am sorry that as far as I am aware this antibody has not specifically been tested for myeneric neurons and we have no data. However, we do guarantee it for rat and mouse samples.

2 Again, PGP9.5 is a neuronal maker and soshould be a good marker for myeneric neurons. However, this has not been specifically tested. Therefore, as both antibodies have not been usedfor specificdetection of myeneric neurons Iam regrettablyunable to say which would be better.

I can suggest the customer could consider trying a further literature search for further information on which of these proteins is the better marker for myeneric neurons.

I am sorry we have no further data to confirm answers to your questions on this occasion. However, i hope this will be helpful. If you have any further questions, please do not hesitate to contact us.

Email: nwaly@unmc.edu

Wht should I use to reconstiute/ Dilute this antibody with? thank you,

ANSWER:

Thank you for your query. As far as I am aware, this antibody is supplied as 25ul of antibody at 19mg/ml so you should not need to reconstitute it.

For future reference, for storage, you can dilute most antibodies in distilled water if they are provided lyophilised in the correct buffer salts and preservatives as this one is (stated on the data sheets).

When you dilute the antibody for your experiments, you will need to dilute them in PBS. Some researchers add 3% BSA to this and a detergent such as tween 20, if they are expecting or having problems with non-specific binding.

I hope this helps. Do please get back to me if you have any more queries and I will be happy to help.

With thanks.

Has the ab10588 been tested for immunofluorescence staining? If so, do you have a protocol for it? I followed a basic protocol and tested with goat serum, BSA, and skim milk blocks so far. I also have some Triton in the buffer. We'll be trying the dilution suggested in your website (10,000x) and incubation at 4C overnight. If you have any particular suggestions with this antibody, please let me know. Also, have you compared ab10588 and ab5392 on histology staining? If so, could you please let me know whether they worked about the same extent and, again, which protocol was followed? Thanks!

ANSWER:

Ab10588 has been tested for immunofluorescence in cultured rat cortical neurons, at the recommended dilution of 1:10,000. I think your blocking stages are good and the incubation overnight should be sufficient for antibody binding to the epitope. My gut feeling is that you may need to use the antibody more concentrated, as you are doing tissue immunofluorescence rather than cell culture immuno. Have you tried a 1:1000 dilution, with similar conditions (i.e extensive PBS wash of your slides, then serum blocking, then the antibody at 1:1000 at 4C overnight in a humid environment)? We have not compared Ab10588 and Ab5392 I'm afraid.

PRE-SALES ENQUIRY Since this interacts with elements of the cytoskeleton, is it specific for neurons among neuroglia, or will it also interact with the neuroglia.

Thanks

ANSWER:

Thank you for your enquiry. MAP2 is only expressed in neurons and not in neuroglia, so the antibody is specific only for neurons. MAP2 antibodies are therefore often used as markers of neuronal lineage cells. If you need any further assistance please do not hesitate to get in touch.

Customer would like to know if the immunogen sequence is available.

ANSWER:

The antibody was raised against full length high molecular weight forms of MAP2 biochemically isolated from cow brain. The full length sequence of bovine high molecular weight MAP2 is not known, but is expected to be very similar to that of human and rodent, which are. A partial cDNA sequence of the bovine MAP2 microtubule binding region can be found at accession S74025. Antibody is polyclonal and therefore probably binds multiple sites down molecule.