Anti-Human BDNF

Product Datasheet

www.leinco.com

excellence in early discovery research

Anti-Human Brain-Derived Neurotrophic Factor (BDNF)

Purified No Carrier Protein

(MGC34632)

Monoclonal Antibody

Prod. No.:	B571
Clone:	35928
Isotype:	Mouse IgG₁
Pkg. Size:	1 mg, 500 µg
Storage:	-20°C to -70°C Detailed storage instructions below.

Description

Specificity:

Mouse Anti-Human Brain-Derived Neurotrophic Factor (BDNF) (Clone 35928) recognizes an epitope on Human BDNF. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype. This antibody shows approximately 5% cross-reactivity with rhβ-NGF and rrβ-NGF

Background:

BDNF is a member of the NGF family of neurotrophic growth factors. Like other members of this family, BDNF supports neuron proliferation and survival. BDNF acts on certain neurons of the central nervous system and the peripheral nervous system, helping to support the survival of existing neurons and encourage the growth and differentiation of new neurons and synapses. In the brain, it is active in the hippocampus, cortex, and basal forebrain—areas vital to learning, memory, and higher thinking. Despite its name, BDNF is actually found in a range of tissue and cell types, not just in the brain. It is also expressed in the retina, the CNS, motor neurons, the kidneys, and the prostate.

BDNF binds at least two receptors on the surface of cells which are capable of responding to this growth factor, TrkB (pronounced "Track B") and the LNGFR (for "low affinity nerve growth factor receptor"). TrkB is a receptor tyrosine kinase (meaning it mediates its actions by causing the addition of phosphate molecules on certain tyrosines in the cell, activating cellular signaling).

Known Species Cross Reactivity: Hu+

Host Species

Mouse

Immunogen

Purified Recombinant Human BDNF (>98%)

Formulation

This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.

Reconstitution

This lyophilized antibody may be aseptically reconstituted with sterile PBS to a final volume of 1 ml. This will result in a final antibody concentration of 500 µg/ml.

Storage and Stability

The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.

Reported Applications

Indirect ELISA, IHC (NBF/Par.), WB

Indirect ELISA: This antibody can be used in an indirect detection ELISA at 0.5 - 1.0 µg/ml with a suitable second step reagent such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114). The detection sensitivity of this indirect ELISA for Human BDNF is approximately 3.0 ng/well.

IHC (NBF/Par.): This antibody should give satisfactory staining results when used at a concentration of 25 µg/ml. The recommended secondary antibody for IHC is Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114). For chromogenic detection with high signal and low background use 3, 3'-Diaminobenzidine Tetrahydrochloride (DAB) Substrate Kit (Leinco Prod. No.: D100) or Optional Metal Enhanced DAB Substrate Kit (Leinco Prod. No.: K107).

Western Blotting: To detect Human BDNF this monoclonal antibody can be used at a concentration of 1-2 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The detection limit for Human BDNF is 25 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. A suitable Western blotting control is Recombinant Human BDNF (Leinco Prod. No.: B106).

Each investigator should determine their own optimal working dilution for specific applications.

Country of Origin

USA