Human NRG1-beta 1/HRG1-beta 1 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 NRG1-beta 1/HRG1-beta 1. 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.
Normal Goat Serum: (Catalog # DY005)
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)
Normal Goat Serum: (Catalog # DY005)
Scientific Data
Product Datasheets
Preparation and Storage
Background: Neuregulin-1 beta 1/NRG1 beta 1
The neuregulin (also known as heregulin) cytokine family is comprised of four genes that encode a large number of secreted or membrane-bound isoforms. All family members share an EGF-like domain that interacts with the ErbB family of tyrosine kinase receptors. The NRG1 gene is the most pleiotropic, encoding more than 14 soluble or transmembrane proteins. NRG1 isoforms can be classified into type I (neu differentiation factor, NDF; heregulin, HRG; acetylcholine receptor inducing activity, ARIA), type II (glial growth factor, GGF) and type III (sensory and motor neuron-derived factor, SMDF) isoforms. NRG proteins show distinct spatial and temporal expression patterns and play important roles during development of both the nervous system and the heart.
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 with NGS, 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 NRG1-beta 1/HRG1-beta 1 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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Anti-inflammatory effects of neuregulin-1 in HaCaT keratinocytes and atopic dermatitis-like mice stimulated with Der p 38
Authors: Yun, JH;Hong, Y;Hong, MH;Kim, G;Lee, JS;Woo, RS;Lee, J;Yang, EJ;Kim, IS;
Cytokine
Species: Human
Sample Types: Cell Culture Supernates
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Engineering Nanofiber Scaffolds with Biomimetic Cues for Differentiation of Skin-Derived Neural Crest-like Stem Cells to Schwann Cells
Authors: AK Podder, MA Mohamed, G Tseropoulo, B Nasiri, ST Andreadis
International Journal of Molecular Sciences, 2022-09-16;23(18):.
Species: Human
Sample Types: Cell Culture Supernates
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Antipsychotic drugs increase Neuregulin1beta1 serum levels in first-episode drug-na�ve patients and chronic schizophrenia with suggestions for improving the treatment of psychotic symptoms
Authors: H Yang, W Pan, W Xiao, M Yang, J Xu, J Li, X Zhang
BMC Psychiatry, 2022-03-25;22(1):217.
Species: Human
Sample Types: Serum
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Oncogenic KRAS promotes growth of lung cancer cells expressing SLC3A2-NRG1 fusion via ADAM17-mediated shedding of NRG1
Authors: DH Shin, SH Kim, M Choi, YK Bae, C Han, BK Choi, SS Kim, JY Han
Oncogene, 2021-11-06;0(0):.
Species: Human
Sample Types: Cell Lysates
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CSF levels of the BACE1 substrate NRG1 correlate with cognition in Alzheimer's disease
Authors: F Mouton-Lig, J Dumurgier, E Cognat, C Hourregue, H Zetterberg, H Vanderstic, E Vanmechele, E Bouaziz-Am, K Blennow, J Hugon, C Paquet
Alzheimers Res Ther, 2020-07-20;12(1):88.
Species: Human
Sample Types: CSF
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Efficient blockade of locally reciprocated tumor-macrophage signaling using a TAM-avid nanotherapy
Authors: SJ Wang, R Li, TSC Ng, G Luthria, MJ Oudin, M Prytyskach, RH Kohler, R Weissleder, DA Lauffenbur, MA Miller
Sci Adv, 2020-05-22;6(21):eaaz8521.
Species: Human
Sample Types: Cell Lysates
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Plasma Fibrin Clot Properties as Determinants of Bleeding Time in Human Subjects: Association with Histidine-Rich Glycoprotein
Authors: K Szu?drzy?s, M Jankowski, DP Potaczek, A Undas
Dis. Markers, 2020-01-25;2020(0):7190828.
Species: Human
Sample Types: Plasma
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Therapeutic efficacy of neuregulin 1-expressing human adipose-derived mesenchymal stem cells for ischemic stroke
Authors: S Ryu, JM Lee, CA Bae, CE Moon, KO Cho
PLoS ONE, 2019-09-27;14(9):e0222587.
Species: Human, Xenograft
Sample Types: Cell Culture Supernates, Tissue Homogenates
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Assessment of a combination of Serum Proteins as potential biomarkers to clinically predict Schizophrenia
Authors: C Li, H Tao, X Yang, X Zhang, Y Liu, Y Tang, A Tang
Int J Med Sci, 2018-06-04;15(9):900-906.
Species: Human
Sample Types: Serum
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Neuregulin 1 discovered as a cleavage target for the HCV NS3/4A protease by a microfluidic membrane protein array
Authors: N Schwartz, M Pellach, Y Glick, R Gil, G Levy, D Avrahami, E Barbiro-Mi, Y Nahmias, D Gerber
N Biotechnol, 2018-02-10;0(0):.
Species: Human
Sample Types: Cell Lysates
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Five-weekly S-1 plus cisplatin therapy combined with trastuzumab therapy in HER2-positive gastric cancer: a phase II trial and biomarker study (WJOG7212G)
Authors: Y Miura, Y Sukawa, S Hironaka, M Mori, K Nishikawa, S Tokunaga, H Okuda, T Sakamoto, K Taku, K Nishikawa, T Moriwaki, Y Negoro, Y Kimura, K Uchino, K Shinozaki, H Shinozaki, N Musha, H Yoshiyama, T Tsuda, Y Miyata, N Sugimoto, T Shirakawa, M Ito, K Yonesaka, K Yoshimura, N Boku, K Nosho, T Takano, I Hyodo
Gastric Cancer, 2017-05-11;0(0):.
Species: Human
Sample Types: Serum
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Patritumab plus trastuzumab and paclitaxel in human epidermal growth factor receptor 2-overexpressing metastatic breast cancer
Cancer Sci., 2016-09-15;107(10):1465-1470.
Species: Human
Sample Types: Serum
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Heregulin-1� and HER3 in hepatocellular carcinoma: status and regulation by insulin
J Exp Clin Cancer Res, 2016-08-11;35(1):126.
Species: Human
Sample Types: Tissue Homogenates
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HER2 phosphorylation is maintained by a PKB negative feedback loop in response to anti-HER2 herceptin in breast cancer.
Authors: Gijsen M, King P, Perera T, Parker PJ, Harris AL, Larijani B, Kong A
PLoS Biol., 2010-12-21;8(12):e1000563.
Species: Human
Sample Types: Cell Lysates
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Neuregulin-ErbB signaling promotes microglial proliferation and chemotaxis contributing to microgliosis and pain after peripheral nerve injury.
Authors: Calvo M, Zhu N, Tsantoulas C, Ma Z, Grist J, Loeb JA, Bennett DL
J. Neurosci., 2010-04-14;30(15):5437-50.
Species: Rat
Sample Types: Tissue Homogenates
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Neuregulin-1, the fetal endothelium, and brain damage in preterm newborns.
Authors: Hoffmann I, Bueter W, Zscheppang K, Brinkhaus MJ, Liese A, Riemke S, Dork T, Dammann O, Dammann CE
Brain Behav. Immun., 2009-09-04;24(5):784-91.
Species: Human
Sample Types: Cell Lysates
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Preventive effects of heregulin-beta1 on macrophage foam cell formation and atherosclerosis.
Authors: Xu G, Watanabe T, Iso Y, Koba S, Sakai T, Nagashima M, Arita S, Hongo S, Ota H, Kobayashi Y, Miyazaki A, Hirano T
Circ. Res., 2009-07-30;105(5):500-10.
Species: Human
Sample Types: Plasma
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Neuregulin-1 beta is associated with disease severity and adverse outcomes in chronic heart failure.
Authors: Ky B, Kimmel SE, Safa RN, Putt ME, Sweitzer NK, Fang JC, Sawyer DB, Cappola TP
Circulation, 2009-07-13;120(4):310-7.
Species: Human
Sample Types: Serum
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Identification of biomarkers in human head and neck tumor cell lines that predict for in vitro sensitivity to gefitinib.
Authors: Hickinson DM, Marshall GB, Beran GJ
Clin Transl Sci, 2009-06-01;2(3):183-92.
Species: Human
Sample Types: Cell Lysates
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Targeting ADAM-mediated ligand cleavage to inhibit HER3 and EGFR pathways in non-small cell lung cancer.
Authors: Zhou BB, Peyton M, He B, Liu C, Girard L, Caudler E, Lo Y, Baribaud F, Mikami I, Reguart N, Yang G, Li Y, Yao W, Vaddi K, Gazdar AF, Friedman SM, Jablons DM, Newton RC, Fridman JS, Minna JD, Scherle PA
Cancer Cell, 2006-07-01;10(1):39-50.
Species: Human
Sample Types: Cell Culture Supernates
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