SpecificityHuman Not yet tested. Antibody may detect mouse and rat proBDNF protein due to high degree of amino acid sequence homology.
Species ReactivityHuman, Mouse (Predicted), Rat
Immunogen DescriptionA synthetic peptide (C-NGPKAGSRGLTS, aa: 47-58) of human proBDNF protein has been used as the immunogen. The sequence is located on the pro-domain of the proBDNF full-length protein.
Application DetailsFlow Cytometry (2 ug/106 cells). Immunocytochemistry (1-2 µg/mL). Other applications not yet tested. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
SpecificityHuman Not yet tested. Antibody may detect mouse and rat proBDNF protein due to high degree of amino acid sequence homology.
Target Host SpeciesHuman
Species ReactivityHuman, Mouse (Predicted), Rat
Antibody HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG1, kappa
Clone NameBS375
ConjugateUnconjugated
Immunogen DescriptionA synthetic peptide (C-NGPKAGSRGLTS, aa: 47-58) of human proBDNF protein has been used as the immunogen. The sequence is located on the pro-domain of the proBDNF full-length protein.
Purity DescriptionProtein G purified mouse IgG
FormatLyophilized from a solution containing PBS buffer pH 7.4 with 3% trehalose, without preservatives.
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 100 µL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material. Final buffer contains no preservatives.
Storage InstructionsAfter reconstitution divide into aliquots and store at -20°C for a higher stability. Antibody contains no preservatives. Store at 2-8°C with an appropriate antibacterial agent. Use sterile methods. Highest purity Glycerol (1:1) may be added for an additional stability when stored at refrigerated or freezing temperatures. Avoid repetitive freeze/thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
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.
Detection of proBDNF expression in human neuroblastoma cell line SHSY-5Y by Flow Cytometry. Fixing and Permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 200 µg/mL Normal Sheep IgG (20 minutes), Primary antibody: Mouse Monoclonal antibody to proBDNF (cat # M-1729-100, 2 μg per ~106 cells) for 30 minutes at room temperature, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1:100 fold dilution) with incubation for 20 minutes in dark at room temperature. Non-specific Control IgG, clone X63 (cat # M-1249-200) was used as negative control under same conditions (black dashed). Flow cytometry data and results were generated using Orflo MoxiflowTM instrument and protocols.
proBDNF staining in fixed and permeabilized rat hippocampal neurons (7 DIV) by Immunocytochemistry. proBDNF-IR (red) is observed in neurites and cell bodies, consistent with expected cellular localization. Blue: Nuclei. Primary antibody concentration: 2 µg/mL; Magnification: 32X. Image courtesy of QBM Cell Science.
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