SpecificityHuman Rat. Antibody is expected to detect mouse proGDNF protein due to 100% amino acid sequence homology.
Species ReactivityHuman, Mouse (Predicted), Rat
Immunogen DescriptionA synthetic peptide (C-EDYPDQFDDVMD, aa: 55-66) of human proGDNF protein has been used as the immunogen. The sequence is located on the pro-domain of the proGDNF full-length protein and is homologous with mouse and rat form of proGDNF.
Product DescriptionMouse anti-Pro-glial cell line-derived neurotrophic factor (proGDNF) Monoclonal Antibody (Unconjugated), suitable for ICC, FC.
Application(s)FC, ICC
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 Rat. Antibody is expected to detect mouse proGDNF protein due to 100% amino acid sequence homology.
Target Host SpeciesHuman
Species ReactivityHuman, Mouse (Predicted), Rat
Antibody HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG3, kappa
Clone NameBS376
ConjugateUnconjugated
Immunogen DescriptionA synthetic peptide (C-EDYPDQFDDVMD, aa: 55-66) of human proGDNF protein has been used as the immunogen. The sequence is located on the pro-domain of the proGDNF full-length protein and is homologous with mouse and rat form of proGDNF.
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 BackgroundNeurotrophic factor that enhances survival and morphological differentiation of dopaminergic neurons and increases their high-affinity dopamine uptake (Ref: uniprot.org). ProGDNF is the unprocessed precursor molecule of mature GDNF and exists as homodimer.
Fixing and Permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 1% BSA, Primary antibody: Mouse Monoclonal antibody to proGDNF (cat # M-1727-100, 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-1249-100) was used as negative control under same conditions (black dashed). Flow cytometry data and results were generated using Orflo MoxiflowTM instrument and protocols. The data demonstrates specific staining of proGDNF expressed in human neuronal cell line SHSY-5Y using cat # M-1727-100.
Immunofluorescence analysis of proGDNF expression in human SH-SY5Y cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) SH-SY5Y cells were incubated with proGDNF antibody M-1727-100 (2 µg/mL, green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 µg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). proGDNF-IR is observed throughout the cytoplasm. Magnification: 100x.
Immunofluorescence analysis of proGDNF expression in rat C6 glioma cells. Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) C6 cells were incubated with proGDNF antibody M-1727-100 (2 µg/mL, green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 µg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). proNGF-IR is observed in the perinuclear cytoplasm. Magnification: 100x.