The Biosensis Mature BDNF/proBDNF Combo RapidTM enzyme-linked immunosorbent assay (ELISA) Kit combines individual, but complementary ELISA kits for the two most important BDNF isoforms: Mature BDNF (BEK-2211) and full-length proBDNF (BEK-2217).

• Complete kit with precise instructions
• Assay duration of less than 4 hours without compromising quality
• Accurate quantification of mature BDNF and proBDNF isoforms in a variety of samples

Both kits are sandwich ELISAs, useful for the quantification of mature BDNF and proBDNF in less than 4 hours in cell culture supernatants, serum, plasma (citrate), cell lysates and brain extract only if used as directed, with a simplified protocol and no loss of sensitivity or specificity. Please refer to the most current kit protocols for BEK-2211 (Mature BDNF RapidTM ELISA) and BEK-2217 (proBDNF RapidTM ELISA), for specific use instructions for each substrate application, in particular blood samples. For human blood samples, we suggest the use of BEK-2241 Mature BDNF/proBDNF Combo kit.

The Mature BDNF ELISA kit consists of a pre-coated mouse monoclonal anti-mature BDNF capture antibody, a biotinylated anti-mature BDNF detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The proBDNF ELISA kit consists of a pre-coated polyclonal anti-proBDNF capture antibody, a biotinylated anti-mature BDNF detection antibody and horseradish peroxidase (HRP)-conjugated streptavidin. The addition of a substrate (3,3',5,5'-tetramethylbenzidine, TMB) yields a coloured reaction product which is directly proportional to the concentration of mature BDNF or proBDNF present in samples and protein standards. A BDNF and proBDNF positive control (QC sample) is provided to assure consistent assay performance.

Both ELISA kits contain high quality protein calibrators. The Mature BDNF ELISA kit employs a recombinant E.coli derived human mature BDNF standard approved by the World Health Organization (WHO, www.nibsc.org). The amino acid sequence of mature BDNF is identical for human, mouse, rat and a number of other species. This kit therefore is suitable to measure mature BDNF in all these species and uses the same antibodies and antigen. The proBDNF ELISA kit employs a recombinant mammalian, cleavage-resistant human proBDNF standard produced by Biosensis and validated against externally available proBDNF proteins. Due to a high degree of amino acid sequence homology, mouse and rat proBDNF can be quantified and expressed as human proBDNF equivalents. Internal Biosensis validation suggests that the use of the human standard provided in this kit will provide estimates that are identical, or close, to the actual levels of rat and mouse proBDNF present in rodent samples.

Mature BDNF ELISA: No cross-reactivity is observed for nerve growth factor (NGF), neurotrophin-3 (NT-3), NT-4/5, glial cell line-derived neurotrophic factor (GDNF) and vascular endothelial growth factor (VEGF165) tested at 25 ng/ml in assay buffer. The reactivity of full-length proBDNF (0.125 ng/mL - 5 ng/ml) was determined in six independent assays using proBDNF proteins from four different sources (mammalian and bacterial, wild-type and mutated). The average cross-reactivity of proBDNF was found to be 5.3% ±0.5% in weight (w/v) concentration, or 12.1% ±1.2% in molar concentration (mean ±SEM).

proBDNF ELISA: A cross-reactivity of 2% in weight concentration (0.9% in molar concentration) has been observed for mature BDNF assayed at 25 ng/ml (893 pmol/l) in Assay Diluent A.Due to a high degree of sequence homology, this human proBDNF ELISA kit cross-reacts with the mouse and rat form of proBDNF. Other species have not yet been tested, but cross-reactivity with a wide range of mammalian forms of proBDNF is expected.The antibodies do not cross-react with nerve growth factor (NGF), neurotrophin-3 (NT-3) or NT-4/5.