Human/Mouse Total COX-2 DuoSet IC 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 IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total COX-2 in cell lysates. An immobilized capture antibody specific for COX-2 binds both phosphorylated and unphosphorylated COX-2. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format.
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
- Available in 2, 5, and 15- (96-well) plate pack sizes
- Economical alternative to Western blot
Kit Content
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
- Streptavidin-HRP
Other Reagents Required
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Lysis Buffer*
IC Diluent*
Blocking Buffer*
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: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent
*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product
Scientific Data
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The Human/Mouse Total COX-2 DuoSet IC ELISA specifically recognizes total COX-2, as shown by Western blot. Lysate prepared from CCD-1070Sk human foreskin fibroblasts treated with 5.0 ng/mL of recombinant human (rh) IL-1 beta (Catalog # 201-LB) for 16 hours was incubated in wells coated with Human/Mouse Total COX-2 Capture Antibody. Unbound material was removed by washing and bound material was solubilized in SDS gel sample buffer. The same lysate and the captured protein were electrophoresed, transferred to a PVDF membrane, and immunoblotted with an Anti-COX-2 Polyclonal Antibody (Catalog # AF4198). Only the band corresponding to human/mouse COX-2 was detected in captured material.
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Quantification of induced COX-2. Human PBMCs, THP-1 human acute monocytic leukemia cells, and RAW 264.7 mouse monocyte/macrophage cells were cultured in the absence or presence of 100 nM Phorbol 12-myristate 13-acetate (PMA) for 48 hours and 1.0 µg/mL of Lipopolysaccharide (LPS) for an additional 24 hours. Human CCD-1070Sk cells were cultured in the absence or presence of 5.0 ng/mL of rhIL-1 beta for 16 hours. The lysates were quantified with this DuoSet IC ELISA. The same lysates were immunoblotted (inset) with an anti-COX-2 polyclonal antibody. The DuoSet IC ELISA results correlate well with the relative amounts of human/mouse COX-2 detected by Western blot.
Product Datasheets
Preparation and Storage
Background: COX-2
Cyclooxygenase (COX), also known as prostaglandin G/H synthase, is a membrane bound enzyme partly responsible for the catalysis of prostanoid synthesis. COX is expressed as at least three different isoforms. COX-1 is constitutively expressed and thought to regulate a number of 'housekeeping' functions such as vascular hemostasis, renal blood flow, and glomerular function. COX-2 expression is tightly regulated and induced by inflammatory mediators such as growth factors, cytokines, and endotoxin. COX-3 appears to be much more strictly regulated spatially and is observed in greatest abundance in cerebral cortex.
Citations for Human/Mouse Total COX-2 DuoSet IC 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.
18
Citations: Showing 1 - 10
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Alpha-linolenic acid protects against methotrexate-induced nephrotoxicity in mouse kidney cells
Authors: Kaplan, HM;Deger, M;Erdogan, KE;Ates, T;Demir, E;
European review for medical and pharmacological sciences
Species: Mouse
Sample Types: Tissue Homogenates
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Comparable Response Following Exposure to Biodiesel and Diesel Exhaust Particles in Advanced Multicellular Human Lung Models
Authors: Rahman, M;Upadhyay, S;Ganguly, K;Introna, M;Ji, J;Boman, C;Muala, A;Blomberg, A;Sandström, T;Palmberg, L;
Toxics
Species: Human
Sample Types: Cell Culture Supernates
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L-Type Ca2+ Channel Inhibition Rescues the LPS-Induced Neuroinflammatory Response and Impairments in Spatial Memory and Dendritic Spine Formation
Authors: J Kim, SG Jeon, HR Jeong, H Park, JI Kim, HS Hoe
International Journal of Molecular Sciences, 2022-11-06;23(21):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Modulation of Inflammatory Responses by a Non-Invasive Physical Plasma Jet during Gingival Wound Healing
Authors: B Eggers, MB Stope, J Marciniak, A Mustea, J Deschner, M Nokhbehsai, FJ Kramer
Cells, 2022-09-02;11(17):.
Species: Human
Sample Types: Cell Lysates
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Nilotinib modulates LPS-induced cognitive impairment and neuroinflammatory responses by regulating P38/STAT3 signaling
Authors: J Kim, HJ Lee, JH Park, BY Cha, HS Hoe
Journal of Neuroinflammation, 2022-07-15;19(1):187.
Species: Mouse
Sample Types: Tissue Homogenates
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Green tea catechins inhibit Porphyromonas gulae LPS-induced inflammatory responses in human gingival epithelial cells: Running title
Authors: S Yoshida, H Inaba, R Nomura, K Nakano, M Matsumoto-
Journal of oral biosciences, 2022-06-02;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Icariin alleviates uveitis by targeting peroxiredoxin 3 to modulate retinal microglia M1/M2 phenotypic polarization
Authors: G Wang, X Li, N Li, X Wang, S He, W Li, W Fan, R Li, J Liu, S Hou
Redox Biology, 2022-03-18;52(0):102297.
Species: Human
Sample Types: Cell Culture Supernates
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Anti-inflammatory effects of Radix aucklandiae herbal preparation ameliorate intestinal mucositis induced by 5-fluorouracil in mice
Authors: JH Liu, CH Hsieh, CY Liu, CW Chang, YJ Chen, TH Tsai
Journal of ethnopharmacology, 2021-02-07;271(0):113912.
Species: Mouse
Sample Types: Tissue Homogenates
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Filifactor alocis and Tumor Necrosis Factor-Alpha Stimulate Synthesis of Visfatin by Human Macrophages
Authors: AVB Nogueira, M Nokhbehsai, A Damanaki, S Eick, C Kirschneck, A Schröder, J Jantsch, J Deschner
International Journal of Molecular Sciences, 2021-01-27;22(3):.
Species: Human
Sample Types: Cell Lysates
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Inhibition of Porphyromonas gulae and periodontal disease in dogs by a combination of clindamycin and interferon alpha
Authors: R Nomura, H Inaba, H Yasuda, M Shirai, Y Kato, M Murakami, N Iwashita, S Shirahata, S Yoshida, S Matayoshi, J Yasuda, N Arai, F Asai, M Matsumoto-, K Nakano
Sci Rep, 2020-02-20;10(1):3113.
Species: Human
Sample Types: Cell Culture Lysates
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Systemic Lipopolysaccharide-Induced Pain Sensitivity and Spinal Inflammation Were Reduced by Minocycline in Neonatal Rats
Authors: CT Hsieh, YJ Lee, X Dai, NB Ojeda, HJ Lee, LT Tien, LW Fan
Int J Mol Sci, 2018-09-27;19(10):.
Species: Rat
Sample Types: Tissue Homogenates
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JIP3 knockout protects mice against high fat diet-induced liver injury
Authors: XJ Ma, HZ Xing, GF Ren, XJ Rao, ZZ Li
Biochem. Biophys. Res. Commun., 2018-02-16;0(0):.
Species: Mouse
Sample Types: Serum
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Honokiol improved chondrogenesis and suppressed inflammation in human umbilical cord derived mesenchymal stem cells via blocking nuclear factor-?B pathway
Authors: H Wu, Z Yin, L Wang, F Li, Y Qiu
BMC Cell Biol., 2017-08-29;18(1):29.
Species: Human
Sample Types: Cell Lysates
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Suppressive Effect of the n-Hexane Extract of Litsea japonica Fruit Flesh on Monosodium-Iodoacetate-Induced Osteoarthritis in Rats
Authors: SH Kim, HJ Choi, WK Yang, JE Lee, JH Cho, IJ Park, S Park, BK Park, M Jin
Evid Based Complement Alternat Med, 2017-08-22;2017(0):1791403.
Species: Rat
Sample Types: Tissue Homogenates
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Anti-inflammatory drugs suppress ultrasound-mediated mesenchymal stromal cell tropism to kidneys
Authors: SR Burks, BA Nguyen, MN Bresler, ME Nagle, SJ Kim, JA Frank
Sci Rep, 2017-08-17;7(1):8607.
Species: Mouse
Sample Types: Tissue Homogenates
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Acupuncture ameliorates inflammatory response in a chronic unpredictable stress rat model of depression
Authors: Jun Lu
Brain Res. Bull, 2016-11-30;0(0):.
Species: Rat
Sample Types: Tissue Homogenates
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Anthocyanins abrogate glutamate-induced AMPK activation, oxidative stress, neuroinflammation, and neurodegeneration in postnatal rat brain
J Neuroinflammation, 2016-11-08;13(1):286.
Species: Mouse
Sample Types: Cell Culture Supernates
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Protective effect of resveratrol against acute lung injury induced by lipopolysaccharide via inhibiting the myd88-dependent Toll-like receptor 4 signaling pathway.
Authors: Zhang Z, Chen N, Liu J, Wu J, Zhang J, Zhang Y, Jiang X
Mol Med Rep, 2014-05-09;10(1):101-6.
Species: Mouse
Sample Types: BALF
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