Nori Human GDNF ELISA Kit Summary

Alternative names for GNF: Glial cell-derived neurotrophic factor

P39905

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: 

Glial cell-derived neurotrophic factor (GDNF) is a protein that is encoded by the GDNF gene.^[1] GDNF is a small protein that potently promotes the survival of many types of neurons and signals through GFRα receptors, particularly GFRα1. It is also responsible for the determination of spermatogonia into primary spermatocytes. GDNF is synthesized as a 211 amino acid-long protein precursor, pro-GDNF^[2] and then cleaved to 134 amino acids.^[2] Proteases that play a role in the proteolysis of pro-GDNF into mature GDNF include furin, PACE4, PC5A, PC5B, and PC7. Because multiple proteases can cleave the pro-GDNF, four different mature forms of GDNF can be produced.^[2] The proteolytic processing of GDNF requires SorLA, a protein sorting receptor. The mature form of the protein is a ligand for the product of the RET protooncogene. GDNF is highly distributed throughout both the peripheral and central nervous system. It can be secreted by astrocytes, oligodendrocytes, Schwann cells, motor neurons, and skeletal muscle during the development and growth of neurons and other peripheral cells.^[2]

The recombinant form of this protein was shown to promote the survival and differentiation of dopaminergic neurons in culture, and was able to prevent apoptosis of motor neurons induced by axotomy. GDNF has the ability to activate the ERK-1 and ERK-2 isoforms of MAP kinase in sympathetic neurons as well as P13K/AKT pathways via activation of its receptor tyrosine kinases.^[3][4] It can also activate Src-family kinases through its GFRα1 receptor.^[5] The most prominent feature of GDNF is its ability to support the survival of dopaminergic^[6] and motor neurons. It prevents apoptosis in motor neurons during development, decreases the overall loss of neurons during development, rescues cells from axotomy-induced death, and prevents chronic degeneration.^[2] GDNF also regulates kidney development and spermatogenesis, and has a powerful and rapid negative (ameliorating) effect on alcohol consumption.^[7] GDNF also promotes hair follicle formation and cutaneous wound healing by targeting resident hair follicle stem cells (BSCs) in the bulge compartment.^[8]

References

1. Lin LF, et al. (1993). Science. 260 (5111): 1130–2.
2. Cintrón-Colón AF, et al. (2020). Cell and Tissue Research. 382 (1): 47–56.
3. Kotzbauer PT, et al. (1996). Nature. 384 (6608): 467–70.
4. Ibáñez CF, Andressoo JO (2017). Neurobiology of Disease. 97 (Pt B): 80–89.
5. Ibáñez CF, Andressoo JO (2017). Neurobiology of Disease. 97 (Pt B): 80–89.
6. Oo TF, et al. (2003). The Journal of Neuroscience. 23 (12): 5141–8.
7. Carnicella S, et al. (2008). Proc Natl Acad Sci USA. 105 (23): 8114–9.
8. Lisse TS, et al. (2020). NPJ Regenerative Medicine. 5 (13): 13.