Alternative NamesPro-brain nerve growth factor; proNGF; NGF
Application(s)WB
Antibody HostMouse
Antibody TypeMonoclonal
SpecificityHuman Species cross-reactivity not tested.
Species ReactivityHuman
Immunogen DescriptionA synthetic peptide (C-HTIPQAHWTKLQ, aa: 30-41) of human proNGF protein has been used as the immunogen. The sequence is located on the pro-domain of the proNGF full-length protein and is 80% homologous to mouse and rat proNGF.
ConjugateBiotin
Purity DescriptionAntibody was purified from cell culture supernatant by Protein G chromatography, biotinylated and buffer-exchanged into PBS, pH 7.4 buffer
Application DetailsThe biotinylated proNGF antibody has been tested by Western Blotting (0.1-0.5 µg/mL) and is also expected to work in applications validated for the unlabelled antibody M-1738-100 at same or higher dilutions: Flow Cytometry and Immunofluorescence.
Other applications not yet tested. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
TargetPro-Nerve growth factor (proNGF)
SpecificityHuman Species cross-reactivity not tested.
Target Host SpeciesHuman
Species ReactivityHuman
Antibody HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG2, lambda
Clone NameBS312
ConjugateBiotin
Immunogen DescriptionA synthetic peptide (C-HTIPQAHWTKLQ, aa: 30-41) of human proNGF protein has been used as the immunogen. The sequence is located on the pro-domain of the proNGF full-length protein and is 80% homologous to mouse and rat proNGF.
Purity DescriptionAntibody was purified from cell culture supernatant by Protein G chromatography, biotinylated and buffer-exchanged into PBS, pH 7.4 buffer
FormatLyophilized from a solution containing PBS buffer pH 7.4 with 3% trehalose, without preservatives.
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 50 µL of sterile-filtered, ultrapure water to give a concentration of 1 mg/mL. Centrifuge to remove any insoluble material. Final buffer contains no preservative.
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).
Alternative NamesPro-brain nerve growth factor; proNGF; NGF
Scientific BackgroundNerve growth factor (NGF) is synthesized as a precursor (proNGF) 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. proNGF is synthesized in target tissues and glia, transported retrogradely and may be released.
Biosensis now offers biotinylated proNGF antibody allowing more flexibility in experimental design by using the biotin-avidin/streptavidin detection method. The ability of biotinylated proNGF antibody to detect proNGF has been validated by WB.
Western blot analysis of proNGF expression in human DU145 prostate cancer cell lysate and normal human brain. Biotinylated, monoclonal proNGF antibody M-1778-50-B (0.5 µg/mL) detects monomeric, non-glycosylated proNGF protein (E.coli derived) at 25 kDa (Lane 1). In DU145 cell lysate (Lane 2) and human brain (Lane 3), M-1778-50-B detects monomeric, glycosylated proNGF at 32 kDa, and several proNGF isoforms (~37 to ~53 kDa). These additional bands have not been characterized, but literature suggests that these bands are at least partly related to glycosylated forms of full-length proNGF (Reinshagen et al., 2000; Lobos et al., 2005; Pedraza et al., 2005; Pundavela et al., 2014). Western Blotting Method: SDS-PAGE: denaturing and reducing, 4-20% Bis-Tris gel; Semi-Dry Transfer: Tris-Glycine (Towbin) buffer with 20% methanol; Membrane: Nitrocellulose (0.45 µm); Blocking: 3% BSA in TBST, 1 hour at RT; Primary antibody: overnight at 4°C; HRP conjugate: streptavidin-HRP (25 ng/mL), 1 hour at RT; Detection: Chemiluminiscence.