The Neurotrophin Antibody Kit contains a collection of sheep antibodies raised against BDNF, NGF, NT3 and NT4/5. The kit is completed with one vial of normal sheep IgG for use as negative control. This kit presents a cost-effective way to investigate neurotrophin protein expression and function by immunological techniques such as immunohistochemistry, ELISA, and inhibition of biological activity.
Neurotrophins are growth factors that regulate neuronal development, maintenance and plasticity. These homo-dimeric proteins activate a multitude of intracellular signalling cascades through their interaction with Trk receptors and p75NTR.
Applications:
Immunohistochemistry: 1-10 µg/ml. BDNF antibody works best on Zamboni's fixed, frozen tissue, and is not recommended for paraffin-embedded tissue.
ELISA: 1-10 µg/ml.
Western Blotting: 1-10 µg/ml. Western blotting is not recommended for the BDNF antibody.
Inhibition of biological activity in vitro/in vivo: Expected ED50 (in vivo) values are in the range of 2-10 µg/ml. The NGF antibody 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. The negative control (normal sheep IgG) should be used at the same concentration as the neurotrophin antibodies.
Specificity:
BDNF antibody: Cross-reactivity of <1% against mouse NGF, recombinant human NT3 or NT4/5 has been shown by ELISA.
NGF antibody: Cross-reactivity of <1% against recombinant human BDNF, NT3 and NT4/5 by ELISA has been shown
NT3 antibody: Cross-reactivity of <1% to mouse NGF, recombinant human BDNF and <5% to NT4/5 has been shown by ELISA.
NT4/5 antibody: <1% cross-reactivity against NGF, recombinant human BDNF and <5% to NT3 has been shown by dot blot.
Reconstitute each vial with sterile-filtered, ultrapure water, to achieve an antibody concentration of 1 mg/ml and centrifuge briefly to remove any insoluble material.
Delivery information: Neurotrophin Antibody Kit (trial size) contains 100 μg sheep IgG of each antibody. Full size kit contains 500 μg sheep IgG of each antibody.
Caution: Note: Store lyophilised, unopened vials at 2 to 8 °C or lower. After reconstitution, prepare aliquots and store at –20 °C for a higher stability. Avoid freeze-thaw cycles.
