Application

ELISA (0.5-1 µg/ml)r>Immunoblotting (1-2 µg/ml)r>Immunohistochemistry (5-15 µg/ml)r>Neutralization Studies (see comments)r>

General description

Anti-BDNF, mouse monoclonal, clone 35928.11, recognizes the ~20 kDa BDNF protein in brain extracts. It is validated for ELISA, Western blotting, immunohistochemistry, and neutralization studies.

Immunoaffinty purified mouse monoclonal antibody. Recognizes the ~20 kDa BDNF protein.

Recognizes the ~20 kDa BDNF protein in brain extracts. Clone 35928.11 was selected based on its ability to neutralize the biological activity of human recombinant BDNF.

Immunogen

recombinant protein consisting of amino acids 129-247 of human BDNF

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Riccio, A., et al. 1997. Science277 1097.Sendtner, M., et al. 1996. Neurochem. Res.21, 831.Barbacid, M. 1995. Ann. N.Y. Acad. Sci.766, 442.Gotz, R., et al. 1994. Nature372, 266.Snider, W.D. 1994. Cell77, 627.Snider, W.D., et al. 1992. J. Neurobiol.23, 1231.

The full-length sequence for BDNF can be found under accession number NP_001700. This antibody has been selected for its ability to neutralize the biological activity of human recombinant BDNF. The exact concentration of antibody required to neutralize human recombinant BDNF activity is dependent on the cytokine concentration, cell type, growth conditions, and the type of activity studied. Suggested neutralization concentration required to yield one-half maximal inhibition of BDNF activity is approximately 5-15 µg/ml in the presence of 2.5 ng/ml of human recombinant BDNF, in a neuron survival assay using embryonic chick dorsal root ganglia neurons. The detection limit for human recombinant BDNF is ~300 ng/lane under non-reducing and reducing conditions. The detection for human recombinant BDNF is ~12.5 ng/ml. Based on immunoblotting and ELISA, this antibody exhibits less than 2% cross-reactivity with human recombinant β-NGF, human recombinant NT-3, and human recombinant NT-4. Antibody should be titrated for optimal results in individual systems.

Packaging

100 µg in Plastic ampoule

Physical form

Lyophilized from PBS, 5% trehalose.

Reconstitution

We recommend resuspending the lyophilized antibody with sterile PBS, pH 7.4, or sterile 20 mM Tris-saline (20 mM Tris containing 0.15 M NaCl), pH 7.4, to yield a final concentration of 100 µg/ml. Following reconstitution, aliquot and freeze (-20°C).

Warning

Toxicity: Standard Handling (A)