We are trying to measure by ELISA (by different distributors) salivary BDNF and milk BDNF without having good results. There is a strong varianilty often the concentrations were under the sensibility (also in pg). Have somebody any suggestions ? Many thanks
Hi Monica, what kit are you using? I used the R&D system and got good results from BDNF in gingival tissue and serum.
Thank you very much for reply. I used kit by Promega (EMax), Cloud clone, Biosensis, also specie-spacific, with or without acidification...I really do not know what I have to do more...
Different years ago I tryed to measure sexual female hormones in serum of female of ass but we observed higher variablity of results. We used your brand type of kit. We asked also to the manufacturing company without any results.
I would like to recommend the RP-HPLC method. Brain-derived neurotrophic factor (BDNF) has Mr 13511.5 and alkaline pI of 9.59, which is very similar to lysozyme (file; RP-HPLC lysozyme). This RP-HPLC method is the reliable protein determination method.
Saliva and milk has many mucin-type large proteins. Therefore, acid extraction pretreatment for sample injection may be good.
Thanks to all of you. Kou, it only remain to find an expertise in RP-HPLC method...that is not in our skills. Thank you again for suggestion
Try http://www.novateinbio.com/en/human-elisa-kits/79974-human-bdnf-elisa-kit.html.
If the assay range is not sufficient, try developing an immuno-PCR, which will increase the sensitivity by atleast 100 fold.
Hi Hope you already found a solution. Otherwise, there are some publications about the use of BDNF ELISA for the milk and saliva samples that might help.
For the milk samples- R&D Systems (human)
Citation: S100B Protein, Brain-Derived Neurotrophic Factor, and Glial Cell Line-Derived Neurotrophic Factor in Human Milk (europepmc.org/articles/PMC3124553)
For saliva- R&D Systems, Insight Genomics or an optimized ELISA (see below)
Citations:
R&D Systems- Higher TNF-α, IGF-1, and Leptin Levels are Found in Tasters than Non-Tasters (europepmc.org/articles/PMC4114300)
Insight Genomics- Transient activation of Hedgehog pathway rescued irradiation-induced hyposalivation by preserving salivary stem/progenitor cells and parasympathetic innervations (europepmc.org/articles/PMC3951215)
Alternatively there is a publication about the optimized salivary BDNF ELISA using monoclonal mouse anti-human BDNF (Calbiochem) europepmc.org/articles/PMC3046426
Their experimental conditions may also be useful in troubleshooting.
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