Human beta-NGF DuoSet ELISA
Human beta-NGF DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human beta-NGF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
Background: beta-NGF
Nerve growth factor (NGF) was initially isolated in the mouse submandibular gland over three decades ago as a 7S complex composed of three non- covalently linked subunits, alpha, beta, and gamma. It is now known that both the alpha and gamma subunits of NGF are members of the kallikrein family of serine proteases, while the beta subunit, called beta-NGF or 2.5S NGF, exhibits all the biological activities ascribed to NGF. The human protein shares approximately 90% identity at the amino acid level with both mouse and rat beta-NGF.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human beta-NGF DuoSet ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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Serum nerve growth factor in horses with osteoarthritis-associated lameness
Authors: A Kendall, C Lützelschw, J Lundblad, E Skiöldebra
Journal of veterinary internal medicine, 2023-04-21;0(0):.
Species: Equine
Sample Types: Serum
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Circulating and Salivary NGF and BDNF Levels in SARS-CoV-2 Infection: Potential Predictor Biomarkers of COVID-19 Disease-Preliminary Data
Authors: F Biamonte, A Re, BO Balzamino, G Ciasca, D Santucci, C Napodano, G Nocca, A Fiorita, M Marino, U Basile, A Micera, CAM Callà
Journal of personalized medicine, 2022-11-09;12(11):.
Species: Human
Sample Types: Serum
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Microglia Impairs Proliferation and Induces Senescence In-Vitro in NGF Releasing Cells Used in Encapsulated Cell Biodelivery for Alzheimer's Disease Therapy
Authors: S Mitra, R Gera, J Sundheimer, M Lemee, LU Wahlberg, B Linderoth, M Eriksdotte, H Behbahani
International Journal of Molecular Sciences, 2022-08-12;23(16):.
Species: Human
Sample Types: Cell Culture Supernates
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Human Primary Astrocytes Differently Respond to Pro- and Anti-Inflammatory Stimuli
Authors: P Szpakowski, D Ksiazek-Wi, M Turniak-Ku, I Pacan, A Glabinski
Biomedicines, 2022-07-22;10(8):.
Species: Human
Sample Types: Cell Culture Supernates
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Amyloid-Beta Peptides and Activated Astroglia Impairs Proliferation of Nerve Growth Factor Releasing Cells In Vitro: Implication for Encapsulated Cell Biodelivery-Mediated AD Therapy
Authors: S Mitra, S Turchetto, W Van Os, LU Wahlberg, B Linderoth, H Behbahani, M Eriksdotte
Cells, 2021-10-21;10(11):.
Species: Human
Sample Types: Cell Culture Supernates
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Peripheral Nerve-Derived Stem Cell Spheroids Induce Functional Recovery and Repair after Spinal Cord Injury in Rodents
Authors: HL Lee, CE Yeum, H Lee, J Oh, JT Kim, WJ Lee, Y Ha, YI Yang, KN Kim
International Journal of Molecular Sciences, 2021-04-16;22(8):.
Species: Human
Sample Types: Cell Culture Supernates
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Influence of Previous COVID-19 and Mastitis Infections on the Secretion of Brain-Derived Neurotrophic Factor and Nerve Growth Factor in Human Milk
Authors: V Demers-Mat, DJ Hines, RM Hines, S Lavangnana, S Fels, E Medo
International Journal of Molecular Sciences, 2021-04-08;22(8):.
Species: Human
Sample Types: Milk Supernates
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Comparison of tear proteomic and neuromediator profiles changes between small incision lenticule extraction (SMILE) and femtosecond laser-assisted in-situ keratomileusis (LASIK)
Authors: YC Liu, GH Yam, MT Lin, E Teo, SK Koh, L Deng, L Zhou, L Tong, JS Mehta
Journal of advanced research, 2020-11-05;29(0):67-81.
Species: Human
Sample Types: Tear Supernates
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Circulating biomarkers and outcomes from a randomised phase 2 trial of gemcitabine versus capecitabine-based chemoradiotherapy for pancreatic cancer
Authors: F Willenbroc, CM Cox, EE Parkes, CS Wilhelm-Be, AG Abraham, R Owens, A Sabbagh, CM Jones, DLI Hughes, T Maughan, CN Hurt, EE O'Neill, S Mukherjee
Br J Cancer, 2020-10-26;0(0):.
Species: Human
Sample Types: Plasma
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NGF from pancreatic stellate cells induces pancreatic cancer proliferation and invasion by PI3K/AKT/GSK signal pathway
Authors: J Jiang, J Bai, T Qin, Z Wang, L Han
J. Cell. Mol. Med., 2020-04-15;0(0):.
Species: Human
Sample Types: cell culture supernatant
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Neuroinflammatory Markers in the Serum of Prepubertal Children with Down Syndrome
Authors: L Tarani, V Carito, G Ferraguti, C Petrella, A Greco, M Ralli, MP Messina, D Rasio, E De Luca, C Putotto, P Versacci, M Ceccanti, M Fiore
J Immunol Res, 2020-03-23;2020(0):6937154.
Species: Human
Sample Types: Serum
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Mast Cells Populate the Corneoscleral Limbus: New Insights for Our Understanding of Limbal Microenvironment
Authors: A Micera, K Jirsova, G Esposito, BO Balzamino, A Di Zazzo, S Bonini
Invest. Ophthalmol. Vis. Sci., 2020-03-09;61(3):43.
Species: Human
Sample Types: Cell Culture Supernates
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Short Communication: Interaction of Nerve Growth Factor (NGF) and Vascular Endothelial Growth Factor (VEGF) in Healthy Individuals
Authors: A Groh, K Jahn, M Gröschl, T Hillemache, J Kornhuber, S Bleich, H Frieling, A Heberlein
Dis. Markers, 2019-11-11;2019(0):7510315.
Species: Human
Sample Types: Serum
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Transforming growth factor-? stimulates nerve growth factor production in osteoarthritic synovium
Authors: S Takano, K Uchida, M Itakura, D Iwase, J Aikawa, G Inoue, M Mukai, M Miyagi, K Murata, H Sekiguchi, M Takaso
BMC Musculoskelet Disord, 2019-05-10;20(1):204.
Species: Human
Sample Types: Cell Culture Supernates
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[EXPRESS] A third HSAN5 mutation disrupts the Nerve Growth Factor furin cleavage site
Authors: S Shaikh, M Nahorski, G Woods
Mol Pain, 2018-10-08;0(0):1744806918809.
Species: Human
Sample Types: Cell Lysates
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Diagnostic Significance of Serum Levels of Nerve Growth Factor and Brain Derived Neurotrophic Factor in Diabetic Peripheral Neuropathy
Authors: Q Sun, DD Tang, EG Yin, LL Wei, P Chen, SP Deng, LL Tu
Med. Sci. Monit., 2018-08-26;24(0):5943-5950.
Species: Human
Sample Types: Serum
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New insights on a NGF-mediated pathway to induce ovulation in rabbits (Oryctolagus cuniculus)
Authors: M Maranesi, L Petrucci, L Leonardi, F Piro, PG Rebollar, P Millán, P Cocci, C Vullo, F Parillo, A Moura, GG Mariscal, C Boiti, M Zerani
Biol. Reprod., 2018-05-01;0(0):.
Species: Rabbit
Sample Types: Plasma
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Excessive nerve growth factor impairs bidirectional communication between the oocyte and cumulus cells resulting in reduced oocyte competence
Authors: Y Zhai, G Yao, F Rao, Y Wang, X Song, F Sun
Reprod. Biol. Endocrinol., 2018-03-27;16(1):28.
Species: Human
Sample Types: Follicular Fluid
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Serum nerve growth factor beta, brain- and glial-derived neurotrophic factor levels and psychopathology in unmedicated patients with schizophrenia
Authors: CS Chu, CL Chu, CC Wu, T Lu
J Chin Med Assoc, 2018-02-01;0(0):.
Species: Human
Sample Types: Serum
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Presence of IL-17 in synovial fluid identifies a potential inflammatory osteoarthritic phenotype
Authors: SJ Snelling, S Bas, GJ Puskas, SG Dakin, D Suva, A Finckh, C Gabay, P Hoffmeyer, AJ Carr, A Lübbeke
PLoS ONE, 2017-04-11;12(4):e0175109.
Species: Human
Sample Types: Serum
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The effects of exogenous lipid on THP-1 cells: an in vitro model of airway aspiration?
Authors: YA Hayman, LR Sadofsky, JD Williamson, SP Hart, AH Morice
ERJ Open Res, 2017-03-20;3(1):.
Species: Human
Sample Types: Cell Culture Supernates
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Equol, a Dietary Daidzein Gut Metabolite Attenuates Microglial Activation and Potentiates Neuroprotection In Vitro
Authors: L Subedi, E Ji, D Shin, J Jin, JH Yeo, SY Kim
Nutrients, 2017-02-27;9(3):.
Species: Human
Sample Types: Cell Culture Supernates
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Neurotrophic tyrosine kinase receptor 1 is a direct transcriptional and epigenetic target of IL-13 involved in allergic inflammation.
Authors: Rochman M, Kartashov A, Caldwell J, Collins M, Stucke E, Kc K, Sherrill J, Herren J, Barski A, Rothenberg M
Mucosal Immunol, 2014-11-12;8(4):785-98.
Species: Human
Sample Types: Cell Lysates
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Over-expression of hNGF in adult human olfactory bulb neural stem cells promotes cell growth and oligodendrocytic differentiation.
Authors: Marei H, Althani A, Afifi N, Abd-Elmaksoud A, Bernardini C, Michetti F, Barba M, Pescatori M, Maira G, Paldino E, Manni L, Casalbore P, Cenciarelli C
PLoS ONE, 2013-12-19;8(12):e82206.
Species: Human
Sample Types: Cell Culture Supernates
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Transplantation of human adipose tissue-derived stem cells delays clinical onset and prolongs life span in ALS mouse model.
Authors: Kim, Kwang S, Lee, Hong J, An, Jin, Kim, Yun B, Ra, Jung Cha, Lim, Inja, Kim, Seung U
Cell Transplant, 2013-09-18;23(12):1585-97.
Species: Human
Sample Types: Cell Culture Supernates
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Long-term delivery of nerve growth factor by encapsulated cell biodelivery in the Gottingen minipig basal forebrain.
Authors: Fjord-Larsen L, Kusk P, Tornoe J, Juliusson B, Torp M, Bjarkam CR, Nielsen MS, Handberg A, Sorensen JC, Wahlberg LU
Mol. Ther., 2010-07-27;18(12):2164-72.
Species: Human
Sample Types: Tissue Homogenates
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Improvement in disability after alemtuzumab treatment of multiple sclerosis is associated with neuroprotective autoimmunity.
Authors: Jones JL, Anderson JM, Phuah CL, Fox EJ, Selmaj K, Margolin D, Lake SL, Palmer J, Thompson SJ, Wilkins A, Webber DJ, Compston DA, Coles AJ
Brain, 2010-07-21;133(0):2232-47.
Species: Human
Sample Types: Cell Culture Supernates
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Nerve growth factor and receptor expression in rheumatoid arthritis and spondyloarthritis.
Authors: Barthel C, Yeremenko N, Jacobs R
Arthritis Res. Ther., 2009-06-02;11(3):R82.
Species: Human
Sample Types: Cell Culture Supernates
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Helminth infections associated with multiple sclerosis induce regulatory B cells.
Authors: Correale J, Farez M, Razzitte G
Ann. Neurol., 2008-08-01;64(2):187-99.
Species: Human
Sample Types: Cell Culture Supernates
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The role of nerve growth factor in hyperosmolar stress induced apoptosis.
Authors: Chang EJ, Im YS, Kay EP
J. Cell. Physiol., 2008-07-01;216(1):69-77.
Species: Human
Sample Types: Cell Culture Supernates
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Neurotrophins modulate monocyte chemotaxis without affecting macrophage function.
Authors: Samah B, Porcheray F, Gras G
Clin. Exp. Immunol., 2008-01-08;151(3):476-86.
Species: Human
Sample Types: Cell Culture Supernates
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Neuroimmune crosstalk in asthma: dual role of the neurotrophin receptor p75NTR.
Authors: Nassenstein C, Kammertoens T, Veres TZ, Uckert W, Spies E, Fuchs B, Krug N, Braun A
J. Allergy Clin. Immunol., 2007-08-22;120(5):1089-96.
Species: Mouse
Sample Types: BALF
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Relationship of serum brain-derived neurotrophic factor level with other markers of disease severity in patients with atopic dermatitis.
Authors: Namura K, Hasegawa G, Egawa M, Matsumoto T, Kobayashi R, Yano T, Katoh N, Kishimoto S, Ohta M, Obayashi H, Ose H, Fukui M, Nakamura N, Yoshikawa T
Clin. Immunol., 2006-12-11;122(2):181-6.
Species: Human
Sample Types: Serum
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Circulating levels of brain-derived neurotrophic factor correlate with disease severity in the intrinsic type of atopic dermatitis.
Authors: Raap U, Werfel T, Goltz C, Deneka N, Langer K, Bruder M, Kapp A, Schmid-Ott G, Wedi B
Allergy, 2006-12-01;61(12):1416-8.
Species: Human
Sample Types: Serum
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Elicitation of immunity to HIV type 1 Gag is determined by Gag structure.
Authors: Young KR, Ross TM
AIDS Res. Hum. Retroviruses, 2006-01-01;22(1):99-108.
Species: Human
Sample Types: Cell Culture Supernates
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Nerve growth factor expression by PLG-mediated lipofection.
Authors: Whittlesey KJ, Shea LD
Biomaterials, 2005-11-28;27(11):2477-86.
Species: Rat
Sample Types: Cell Culture Supernates
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Neurotrophic factors in relapsing remitting and secondary progressive multiple sclerosis patients during interferon beta therapy.
Authors: Caggiula M, Batocchi AP, Frisullo G, Angelucci F, Patanella AK, Sancricca C, Nociti V, Tonali PA, Mirabella M
Clin. Immunol., 2005-11-07;118(1):77-82.
Species: Human
Sample Types: Cell Culture Supernates
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Changes in levels of nerve growth factor in nasal secretions after capsaicin inhalation in patients with airway symptoms from scents and chemicals.
Authors: Millqvist E, Ternesten-Hasseus E, Stahl A, Bende M
Environ. Health Perspect., 2005-07-01;113(7):849-52.
Species: Human
Sample Types: Nasal Lavage Fluid
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Used for measuring NGF concentrations in bovine seminal plasma. Worked very well and instructions were easy to follow.