Alternative NamesProform brain derived neurotrophic factor
Application(s)ELISA, Neutralize, WB
Antibody HostSheep
Antibody TypePolyclonal
SpecificityConfirmed to react with purified human proBDNF, crossreact with mouse and rat proBDNF Cross reactivity with other species than human, mouse and rat has not yet been tested
Species ReactivityHuman, Mouse, Rat
Immunogen DescriptionThe recombinant prodomain fragment of human brain-derived neurotrophic factor
Application DetailsELISA, Western Blot, biological neutralization of proBDNF, Immunocytochemistry/Immunofluorescence. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
SpecificityConfirmed to react with purified human proBDNF, crossreact with mouse and rat proBDNF Cross reactivity with other species than human, mouse and rat has not yet been tested
Target Host SpeciesHuman
Species ReactivityHuman, Mouse, Rat
Antibody HostSheep
Antibody TypePolyclonal
Antibody IsotypeMixed
ConjugateUnconjugated
Immunogen DescriptionThe recombinant prodomain fragment of human brain-derived neurotrophic factor
Purity DescriptionWhole Serum
FormatLyophilized
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 100 µL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material.
Storage InstructionsAfter reconstitution keep aliquots at -20ºC for a higher stability, and at 2-8°C with an appropriate antibacterial agent. Glycerol (1:1) may be added for an additional stability. Avoid repetitive freeze/thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Alternative NamesProform brain derived neurotrophic factor
Scientific BackgroundBrain derived neurotrophic factor (BDNF) is synthesized as a precursor (proBDNF) which may be released and have physiological functions to cause cell death. It binds neurotrophin receptor p75 and sortilin and may also be important for the development of nervous system. proBDNF is synthesized in neurons and glia (eg., microglia), transported anterogradely and retrogradely and may be released in an activity dependent manner. This antibody is raised in sheep to detect the prodomain of BDNF and not the mature peptide.
Western blot analysis of proBDNF on DRG homogenate. lane 1: brain; lane 2: DRG. The antibody recognises monomer and dimer of the recombinant proBDNF.
Immunofluorescence analysis of proBDNF expression in human SHSY-5Y cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) SHSY-5Y cells were incubated with proBDNF antibody S-079-100 (1:1000, red) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-sheep-PE antibody (1:100, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). BDNF-IR is observed in the perinuclear cytoplasm. Magnification: 100x.
Separation and detection of proBDNF in conditioned 293 cell culture medium (Lanes 1 and 3, 50 ng/lane) and purified mature BDNF (Lanes 2 and 4, 50 ng/lane) by reduced SDS-PAGE and western blotting. Anti-proBDNF sheep antibody S-079-100 (1/1000) detects 2 bands (Lane 1) with the apparent molecular weight of ≈ 32 kDa (red arrow) and ≈ 64 kDa (blue arrow) corresponding to glycosylated proBDNF monomer and dimer, respectively. S-079-100 is specific for proBDNF and does not recognize mature BDNF (Lane 2). Rabbit anti-BDNF antibody R-1707-100 (0.2 µg/mL) is a superb antibody for western blotting. It detects mature BDNF (≈ 14 kDa, Lane 4, black arrow) and shows lower affinity for pro-BDNF monomer (Lane 3, red arrow) and proBDNF dimer (Lane 3, blue arrow).