SpecificityA cross reactivity of less than 1% to recombinant human BDNF, NT3, NT4/5 by ELISA has been shown. This antiserum is known to cross react with mouse, rat, human and avian NGF bot not bovine NGF.
Species ReactivityAvian, Human, Mouse, Rat
Immunogen DescriptionNative mouse beta NGF purified from submaxillary salivary gland (95% purity by PAGE)
Application DetailsIHC, 1-site ELISA, WB, immunoblot, inhibition of biological activity. A dilution of 1:1000-1:5000 is recommended for IHC, western blot and immunoblot; 1:15000 for ELISA; for inhibition of biological activity: 1:10-50 for in vitro, 5-10 µL/g body weight for in vivo. This antiserum completely inhibits neuronal survival and the outgrowth actions of murine NGF in chicken DRG in vitro. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
TargetBeta-nerve growth factor (beta-NGF)
SpecificityA cross reactivity of less than 1% to recombinant human BDNF, NT3, NT4/5 by ELISA has been shown. This antiserum is known to cross react with mouse, rat, human and avian NGF bot not bovine NGF.
Target Host SpeciesMouse
Species ReactivityAvian, Human, Mouse, Rat
Antibody HostRabbit
Antibody TypePolyclonal
Antibody IsotypeMixed
ConjugateUnconjugated
Immunogen DescriptionNative mouse beta NGF purified from submaxillary salivary gland (95% purity by PAGE)
Purity DescriptionWhole serum
FormatLyophilized
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 100 µL of sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material.
Storage InstructionsStore lyophilized antibody at 2-8ºC. After reconstitution keep aliquots at -20°C to -80ºC for a higher stability, and at 2-8°C with an appropriate antibacterial agent. Avoid repetitive freeze/thaw cycles. Glycerol (1:1) may be added for an additional stability.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Scientific BackgroundFUNCTION: Nerve growth factor is important for the development and maintenance of the sympathetic and sensory nervous systems. It stimulates division and differentiation of sympathetic and embryonic sensory neurons. SUBUNIT: Homodimer, associated by noncovalent forces. SUBCELLULAR LOCATION: Secreted protein. SIMILARITY: Belongs to the NGF-beta family.
Western Blot analysis of NGF expression in rodent and human homogenates. Mature NGF is detected in mouse salivary gland homogenate at ~13 kDa, while tissue samples reveal predominantly proNGF isoforms at higher molecular weights. This is consistent with the observation that proNGF is the predominant isoform in brain tissue (Fahnenstock et al., 2001). Several proNGF-bands are observed, likely representing differently glycosylated proNGF-isoforms as reported by Reinshagen et al., 2000, Lobos et al., 2005, Pedraza et al., 2005 and Pundavela et al., 2014. Incubation of membrane with pre-immune serum reveals some non-specific bands at ~40 kDa, while overall confirming NGF-immunoreactivity.
Western Blotting Method: SDS-PAGE: denaturing and reducing, 10% Bis-Tris gel; Semi-Dry Transfer: Tris-Glycine (Towbins) buffer with 20% methanol; Membrane: PVDF (0.45 µm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: 5 µg/mL, overnight at 2-8°C; Secondary antibody: anti-rabbit-HRP (1/20,000), 2 hours at RT; Detection: Chemiluminiscence.
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