Description

Nori Human Pro-BDNF ELISA Kit Summary

Alternative names for Pro-BDNF: Pro- Brain-derived neurotrophic factor

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Brain-derived neurotrophic factor, also known as BDNF, is a secreted protein^[1] that is encoded by the BDNF gene.^[2] BDNF is a member of the “neurotrophin” family of growth factors, which are related to the canonical “Nerve Growth Factor”, NGF. Neurotrophic factors are found in the brain and the periphery. 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.^[3] In the brain, it is active in the hippocampus, cortex, and basal forebrain—areas vital to learning, memory, and higher thinking.^[4] BDNF itself is important for long-term memory.^[5] BDNF was the second neurotrophic factor to be characterized after nerve growth factor (NGF). BDNF is made in the endoplasmic reticulum and secreted from dense-core vesicles. It binds carboxypeptidase E (CPE), and the disruption of this binding has been proposed to cause the loss of sorting of BDNF into dense-core vesicles. The phenotype for BDNF knockout mice can be severe, including postnatal lethality. Other traits include sensory neuron losses that affect coordination, balance, hearing, taste, and breathing. Knockout mice also exhibit cerebellar abnormalities and an increase in the number of sympathetic neurons. Various studies have shown possible links between BDNF and conditions such as depression,^[6][7] bipolar disorder,^[8] schizophrenia, obsessive-compulsive disorder, Alzheimer’s disease, Huntington’s disease, Rett syndrome, and dementia, as well as anorexia nervosa and bulimia nervosa. Like all neurotrophins, BDNF is initially synthesized as a precursor (proBDNF), which is subsequently cleaved to generate mature (m)BDNF. proBDNF interacts preferentially with the pan-neurotrophin receptor p75 (p75^NTR), whereas mBDNF selectively binds and activates the receptor tyrosine kinase TrkB.^[9]

References

1. Robinson RC, Radziejewski C, Stuart DI, Jones EY (April 1995). Biochemistry 34 (13): 4139–46.
2. Jones KR, Reichardt LF (October 1990). Proc. Natl. Acad. Sci. U.S.A. 87 (20): 8060–4.
3. Acheson A, Conover JC, Fandl JP, et al. (March 1995). Nature 374 (6521): 450–3.
4. Yamada K, Nabeshima T (April 2003). J. Pharmacol. Sci. 91 (4): 267–70.
5. Bekinschtein P, Cammarota M, et al. (2008). Proc. Natl. Acad. Sci. U.S.A. 105 (7): 2711–6.
6. Dwivedi Y (2009). Neuropsychiatr Dis Treat 5: 433–49.
7. Brunoni AR, Lopes M, Fregni F (2008). Int. J. Neuropsychopharmacol. 11 (8): 1169–80.
8. Iria Grande, Gabriel Rodrigo Fries, et al. Psychiatry Investig. 2010 December; 7(4): 243–250.
9. Je HS et al. (2012) Proc Natl Acad Sci USA 109 (39):15924-5.

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