Yes. See for example Giralt et al., 2009, Neuroscience 158:1234-1250 (attached)

Yes, both DNA and protein gels work very well for BDNF and have been established for a long time.

Yes. At contrast with ELISA, it allows the determination of pro- and mature forms of BDNF (see Quirié et al., PlosOne 2012)

Briefly, yes, you can easily use Western-blot to detect BDNF levels from rat/mouse brain extracts. Because you separate your proteins based on their molecular weight, you will be able to discriminate between pro- and truncated forms of BDNF if you use the appropriate antibody. Some of them will be more specific than others to pro-BDNF depending on the epitope, but if you want to look at both, you can try with the Santa Cruz N-20.

I have to say that the sensitivity will not be as good as with an ELISA though, so if you're really interested in an accurate quantification, I would go with the ELISA (some of them have an extraction protocol allowing you to look at only the mature form if that is what you're interested in). If you're expecting big differences and variations between your biological conditions, then the Western-blot should be enough.

Of course, the combination of both would be the best: using ELISA to accurately quantifiy, and confirm the specific form involved by western-blot.

Yes, absolutely. You can easily measure levels of BDNF and pro-BDNF via Western blotting. Let me know if you need suggestions for antibodies.

I have just seen this thread and i am having trouble with BDNF myself. I am currently using a total prep for my lysate (and RIPA buffer) but i am getting nothing. I have also tried nto heating the sample up so much before as well (37 degrees, and leaving at room temp, but neither help. Is there a different tissue prep which is better for BDNF detection? e.g. fractionation??? any help would be great, im totally stuck!

I too saw this thread and wanted to ask even though it is an older one.

I am trying to extract and measure BDNF w/Western blots of mouse hippocampus, cerebellum, and frontal cortex with a 1% SDS, 0.5% Tris-HCL(pH8.0), water and roche tablet as an extraction buffer. I either think I have a problem with the BDNF antibody (Santa Cruz H117, sc-20981; 1:200) or the extraction or samples possibly being too dilute.

The samples were all pretty dilute too begin with, and I somewhat struggled to get even 15ug of protein into the wells with the gel containing hippocampal lysate, however, the cerebellar samples were much more concentrated and fit into the well of that gel easily.

The membranes were fine and GAPDH was very apparent with both tissues. With hippocampus there was a fainter band right below the GAPDH (which I am now assuming is pro-BDNF), but no other bands except some high MW junk. What's stranger though is that the membrane with the cerebellar proteins showed the exact same things except no bands below GAPDH at all. We used 10% pre-made bio-rad gel and 0.2micron Nitrocellulose. Any suggestions would be greatly appreciated! Thank you.