Human beta-NGF DuoSet ELISA

Catalog # Availability Size / Price Qty
DY256
DY256-05
Ancillary Products Available
Human beta-NGF ELISA Standard Curve
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Product Details
Procedure
Citations (33)
FAQs
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Human beta-NGF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 pg/mL
Sufficient Materials
For five or fifteen 96-well plates*
Specificity
Please see the product datasheet

* 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)

Data Example

Human beta-NGF ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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.

Long Name:
beta Nerve Growth Factor
Entrez Gene IDs:
4803 (Human); 18049 (Mouse); 310738 (Rat); 403402 (Canine); 103350089 (Rabbit)
Alternate Names:
Beta nerve growth factor precursor; beta-nerve growth factor; betaNGF; beta-NGF; HSAN5; MGC161426; nerve growth factor (beta polypeptide); nerve growth factor, beta subunit; NGFB; NGFBMGC161428

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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.

33 Citations: Showing 1 - 10
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  1. 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, 2021;29(0):67-81.
    Species: Human
    Sample Types: Tear Supernates
  2. 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;22(8):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. 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;22(8):.
    Species: Human
    Sample Types: Milk Supernates
  4. 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;2020(0):6937154.
    Species: Human
    Sample Types: Serum
  5. 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;0(0):.
    Species: Human
    Sample Types: cell culture supernatant
  6. 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;61(3):43.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. 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;0(0):.
    Species: Human
    Sample Types: Plasma
  8. 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;20(1):204.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. 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;2019(0):7510315.
    Species: Human
    Sample Types: Serum
  10. 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;16(1):28.
    Species: Human
    Sample Types: Follicular Fluid
  11. 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;24(0):5943-5950.
    Species: Human
    Sample Types: Serum
  12. 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;0(0):.
    Species: Rabbit
    Sample Types: Plasma
  13. 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;0(0):.
    Species: Human
    Sample Types: Serum
  14. [EXPRESS] A third HSAN5 mutation disrupts the Nerve Growth Factor furin cleavage site
    Authors: S Shaikh, M Nahorski, G Woods
    Mol Pain, 2018;0(0):1744806918809.
    Species: Human
    Sample Types: Cell Lysates
  15. 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;9(3):.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. 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;3(1):.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. 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;12(4):e0175109.
    Species: Human
    Sample Types: Serum
  18. 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, 2015;8(4):785-98.
    Species: Human
    Sample Types: Cell Lysates
  19. 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, 2014;23(12):1585-97.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. 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;8(12):e82206.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. 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;18(12):2164-72.
  22. 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;133(0):2232-47.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Nerve growth factor and receptor expression in rheumatoid arthritis and spondyloarthritis.
    Authors: Barthel C, Yeremenko N, Jacobs R
    Arthritis Res. Ther., 2009;11(3):R82.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. The role of nerve growth factor in hyperosmolar stress induced apoptosis.
    Authors: Chang EJ, Im YS, Kay EP
    J. Cell. Physiol., 2008;216(1):69-77.
    Species: Human
    Sample Types: Cell Culture Supernates
  25. Neurotrophins modulate monocyte chemotaxis without affecting macrophage function.
    Authors: Samah B, Porcheray F, Gras G
    Clin. Exp. Immunol., 2008;151(3):476-86.
    Species: Human
    Sample Types: Cell Culture Supernates
  26. Helminth infections associated with multiple sclerosis induce regulatory B cells.
    Authors: Correale J, Farez M, Razzitte G
    Ann. Neurol., 2008;64(2):187-99.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. 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;120(5):1089-96.
    Species: Mouse
    Sample Types: BALF
  28. 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., 2007;122(2):181-6.
    Species: Human
    Sample Types: Serum
  29. Elicitation of immunity to HIV type 1 Gag is determined by Gag structure.
    Authors: Young KR, Ross TM
    AIDS Res. Hum. Retroviruses, 2006;22(1):99-108.
    Species: Human
    Sample Types: Cell Culture Supernates
  30. 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;61(12):1416-8.
    Species: Human
    Sample Types: Serum
  31. Nerve growth factor expression by PLG-mediated lipofection.
    Authors: Whittlesey KJ, Shea LD
    Biomaterials, 2006;27(11):2477-86.
    Species: Rat
    Sample Types: Cell Culture Supernates
  32. 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;118(1):77-82.
    Species: Human
    Sample Types: Cell Culture Supernates
  33. 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;113(7):849-52.
    Species: Human
    Sample Types: Nasal Lavage Fluid

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Human beta-NGF DuoSet ELISA
By Anonymous on 05/30/2017
Application: Sample Tested: seminal plasma

Used for measuring NGF concentrations in bovine seminal plasma. Worked very well and instructions were easy to follow.