Western Blotting (denaturing and reducing): 0.2 to 2 µg/mL. M-1744-100 antibody detects 14 kDa mature BDNF monomer in human serum human and acid treated human and rodent whole tissue homogenates. In simple Tris-homogenate human brain lysates, M-1744-100 detects mature BDNF at 14kDa as well as an uncharacterized band at ~50kDa. No proBDNF band is detected in our samples, although M-1744-100 will detect proBDNF in purified form on blots. Additionally, M-1744-100 detects an additional band 18 kDa BDNF isoform human serum. For cell lysates, some caution is advised as in our hands M-1744-100 fails to detect mature BDNF on SH-SY5Y or C6 cell lysates but does detect mature BDNF in neuronal cell culture lysates. The reason for these differences has not been characterized. For detection of mature BDNF in cell lysates such as SH-SY5Y we recommend affinity-purified rabbit polyclonal antibody to rhBDNF (R-1707-100), or rabbit polyclonal antibody to BDNF peptide 1-10 (R-083-100, whole serum; R-066-500, IgG).
Immunofluorescence: 1 to 4 µg/mL, 4% formaldehyde fixation and permeabilization. Staining can be weak until optimized.
M-1744-100 is not recommended for Flow Cytometry at this time.
Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
