Rat beta-NGF DuoSet ELISA

Catalog # Availability Size / Price Qty
DY556
Ancillary Products Available
Rat beta-NGF ELISA Standard Curve
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Product Details
Procedure
Citations (26)
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Rat beta-NGF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
15.6 - 1,000 pg/mL
Sufficient Materials
For 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 rat 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

Rat beta-NGF ELISA Standard Curve

Product Datasheets

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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 Rat 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.

26 Citations: Showing 1 - 10
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  1. Artificial nerve graft constructed by coculture of activated Schwann cells and human hair keratin for repair of peripheral nerve defects
    Authors: HJ Qin, H Li, JZ Chen, KR Zhang, XQ Zhao, JQ Qin, B Yu, J Yang
    Neural regeneration research, 2023;18(5):1118-1123.
    Species: Rat
    Sample Types: Cell Culture Supernates
  2. Stimulation of Neurite Outgrowth Using Autologous NGF Bound at the Surface of a Fibrous Substrate
    Authors: MR Casanova, RL Reis, A Martins, NM Neves
    Biomolecules, 2021;12(1):.
    Species: Rat
    Sample Types: Plasma
  3. TGF-beta regulates nerve growth factor expression in a mouse intervertebral disc injury model
    Authors: Y Yokozeki, K Uchida, A Kawakubo, M Nakawaki, T Okubo, M Miyagi, G Inoue, M Itakura, H Sekiguchi, M Takaso
    Bmc Musculoskeletal Disorders, 2021;22(1):634.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. Dental pulp stem cells stimulate neuronal differentiation of PC12 cells
    Authors: N Sultan, LE Amin, AR Zaher, ME Grawish, BA Scheven
    Neural regeneration research, 2021;16(9):1821-1828.
    Species: Rat
    Sample Types: Cell Culture Supernates
  5. Quercetin protects against diabetic retinopathy in rats by inducing heme oxygenase-1 expression
    Authors: GR Chai, S Liu, HW Yang, XL Chen
    Neural regeneration research, 2021;16(7):1344-1350.
    Species: Rat
    Sample Types: Cell Culture Supernates
  6. Microenvironment-responsive immunoregulatory electrospun fibers for promoting nerve function recovery
    Authors: K Xi, Y Gu, J Tang, H Chen, Y Xu, L Wu, F Cai, L Deng, H Yang, Q Shi, W Cui, L Chen
    Nat Commun, 2020;11(1):4504.
    Species: Rat
    Sample Types: Protein
  7. Nerve growth factor continuously elevates in a rat rotator cuff tear model
    Authors: N Nagura, T Kenmoku, K Uchida, M Nakawaki, G Inoue, M Takaso
    J Shoulder Elbow Surg, 2019;28(1):143-148.
    Species: Rat
    Sample Types: Tissue Homogenates
  8. Insulin treatment protects the brain against neuroinflammation by reducing cerebral cytokines and modulating mitochondrial function
    Authors: P Bortoluzzi, D Casagrande, N Dos Santos, K Ugioni Sim, C Mendes, ME Anastácio, PC Lock Silve, A Pastoris M
    Brain Res. Bull., 2019;0(0):.
    Species: Rat
    Sample Types: Tissue Homogenates
  9. Electroacupuncture and moxibustion promote regeneration of injured sciatic nerve through Schwann cell proliferation and nerve growth factor secretion
    Authors: LN Hu, JX Tian, W Gao, J Zhu, FF Mou, XC Ye, YP Liu, PP Lu, SJ Shao, HD Guo
    Neural Regen Res, 2018;13(3):477-483.
    Species: Rat
    Sample Types: Serum
  10. Lifelong environmental enrichment in the absence of exercise protects the brain from age-related cognitive decline
    Authors: AM Birch, ÁM Kelly
    Neuropharmacology, 2018;0(0):.
    Species: Rat
    Sample Types: Tissue Culture Supernates
  11. Isoflurane and the analgesic effect of acupuncture and electroacupuncture in an animal model of neuropathic pain
    Authors: LN Spezia Ada, R Vercelino, C de Oliveir, VL Scarabelot, A de Souza, LF Medeiros, SG Cioato, W Caumo, ILS Torres
    J Acupunct Meridian Stud, 2018;0(0):.
    Species: Rat
    Sample Types: Tissue Homogenates
  12. Therapeutic Benefit for Late, but Not Early, Passage Mesenchymal Stem Cells on Pain Behaviour in an Animal Model of Osteoarthritis
    Authors: V Chapman, H Markides, DR Sagar, L Xu, JJ Burston, P Mapp, A Kay, RH Morris, O Kehoe, AJ El Haj
    Stem Cells Int, 2017;2017(0):2905104.
    Species: Rat
    Sample Types: Serum
  13. Malathion increases apoptotic cell death by inducing lysosomal membrane permeabilization in N2a neuroblastoma cells: a model for neurodegeneration in Alzheimer's disease
    Authors: R Venkatesan, YU Park, E Ji, EJ Yeo, SY Kim
    Cell Death Discov, 2017;3(0):17007.
    Species: Rat
    Sample Types: Cell Culture Supernates
  14. A biphenyl derivative from the twigs of Chaenomeles speciosa
    Authors: WS Suh, KJ Park, DH Kim, L Subedi, SY Kim, SU Choi, KR Lee
    Bioorg. Chem., 2017;72(0):156-160.
    Species: Rat
    Sample Types: Cell Culture Supernates
  15. Time-Dependent Nerve Growth Factor Signaling Changes in the Rat Retina During Optic Nerve Crush-Induced Degeneration of Retinal Ganglion Cells
    Authors: LA Mesentier-, S De Nicol•, P Rosso, LA De Vitis, V Castoldi, L Leocani, R Mendez-Ote, MF Santiago, P Tirassa, P Rama, A Lambiase
    Int J Mol Sci, 2017;18(1):.
    Species: Rat
    Sample Types: Tissue Homogenates
  16. Incorporation of chitosan microspheres into collagen-chitosan scaffolds for the controlled release of nerve growth factor.
    Authors: Zeng, Wen, Rong, Mengyao, Hu, Xueyu, Xiao, Wei, Qi, Fengyu, Huang, Jinghui, Luo, Zhuojing
    PLoS ONE, 2014;9(7):e101300.
    Species: Rat
    Sample Types: Cell Culture Supernates
  17. Rab7 silencing prevents mu-opioid receptor lysosomal targeting and rescues opioid responsiveness to strengthen diabetic neuropathic pain therapy.
    Authors: Mousa S, Shaqura M, Khalefa B, Zollner C, Schaad L, Schneider J, Shippenberg T, Richter J, Hellweg R, Shakibaei M, Schafer M
    Diabetes, 2013;62(4):1308-19.
    Species: Rat
    Sample Types: Tissue Homogenates
  18. Electrical stimulation of sympathetic neurons induces autocrine/paracrine effects of NGF mediated by TrkA.
    Authors: Saygili E, Schauerte P, Kuppers F, Heck L, Weis J, Weber C, Schwinger RH, Hoffmann R, Schroder JW, Marx N, Rana OR
    J. Mol. Cell. Cardiol., 2010;49(1):79-87.
    Species: Rat
    Sample Types: Cell Culture Supernates
  19. Effects of nicergoline on corneal epithelial wound healing in rat eyes.
    Authors: Kim SY, Choi JS, Joo CK
    Invest. Ophthalmol. Vis. Sci., 2009;50(2):621-5.
    Species: Rat
    Sample Types: Cell Culture Supernates
  20. Low-frequency electro-acupuncture reduces the nociceptive response and the pain mediator enhancement induced by nerve growth factor.
    Authors: Aloe L, Manni L
    Neurosci. Lett., 2009;449(3):173-7.
    Species: Rat
    Sample Types: Serum
  21. Anti-NGF-antibody administration as collyrium reduces the presence of NGF and enhances the expression of VEGF in the retina, lacrimal gland and hippocampus.
    Authors: Colafrancesco V, Cirulli F, Rossi S, Berry A, Aloe L
    Neurosci. Lett., 2009;463(3):203-6.
    Species: Rat
    Sample Types: Tissue Homogenates
  22. The multi-herbal medicine Gongjin-dan enhances memory and learning tasks via NGF regulation.
    Authors: Moon E, Her Y, Lee JB, Park JH, Lee EH, Kim SH, Oh MS, Jang CG, Kim SY
    Neurosci. Lett., 2009;466(3):114-9.
    Species: Rat
    Sample Types: Cell Culture Supernates
  23. Eosinophil-nerve interactions and neuronal plasticity in rat gut associated lymphoid tissue (GALT) in response to enteric parasitism.
    Authors: O'Brien LM, Fitzpatrick E, Baird AW, Campion DP
    J. Neuroimmunol., 2008;197(1):1-9.
    Species: Rat
    Sample Types: Tissue Homogenates
  24. Maternal poly I:C exposure during pregnancy regulates TNF alpha, BDNF, and NGF expression in neonatal brain and the maternal-fetal unit of the rat.
    Authors: Gilmore JH, Jarskog LF, Vadlamudi S
    J. Neuroimmunol., 2004;159(1):106-12.
    Species: Rat
    Sample Types: Tissue Homogenates
  25. Intracerebral transplantation of carotid body in rats with transient middle cerebral artery occlusion.
    Authors: Yu G, Xu L, Hadman M, Hess DC, Borlongan CV
    Brain Res., 2004;1015(1):50-6.
    Species: Rat
    Sample Types: Tissue Homogenates
  26. Intravenous administration of marrow stromal cells (MSCs) increases the expression of growth factors in rat brain after traumatic brain injury.
    Authors: Mahmood A, Lu D, Chopp M
    J. Neurotrauma, 2004;21(1):33-9.
    Species: Rat
    Sample Types: Tissue Homogenates

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