Human/Mouse Total COX-2 DuoSet IC ELISA

Name: Human/Mouse Total COX-2 DuoSet IC ELISA
Brand: R&D Systems
SKU: DYC4198-2

R&D Systems, Inc. a Bio-Techne Brand

Catalog # DYC4198-2

Citations (15)

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Key Product Details

Assay Type:	Solid Phase Sandwich ELISA
Assay Range:	156 - 10,000 pg/mL
Sample Types:	Cell Lysates
Human/Mouse Total COX-2 DuoSet IC ELISA 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

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Summary for Human/Mouse Total COX-2 DuoSet IC ELISA

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 Specifications for Human/Mouse Total COX-2 DuoSet IC ELISA

Assay Format	96-well strip plate (sold separately)
Sample Volume Required	Cell lysates (100 µL)
Reactivity	Human, Mouse
Specificity	Please see the product datasheet
Detection Method	Colorimetric ELISA - 450nm (TMB)
Conjugate	Biotin
Label	HRP
Measurement	Quantitative ELISA
Sufficient Materials	Kits available for two, five, or fifteen 96-well plates*

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

Kit Contents for Human/Mouse Total COX-2 DuoSet IC ELISA

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 Examples for Human/Mouse Total COX-2 DuoSet IC ELISA

	Standard Curve
	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.
	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.

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: 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.

Item	Value
Long Name	Cyclooxygenase 2
Entrez Gene IDs	5743 (Human); 19225 (Mouse); 29527 (Rat)
Alternate Names	COX-2, COX2, COX2cyclooxygenase 2b, EC 1.14.99, EC 1.14.99.1, GRIPGHS, PGG/HS, PGH synthase 2, PGHS-2, PHS II, PHS-2, PHS-II, PTGS2, Prostaglandin H2 synthase 2, Prostaglandin-endoperoxide synthase 2, cyclooxygenase-2, hCox-2, prostaglandin G/H synthase 2, prostaglandin G/H synthase and cyclooxygenase, prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase andcyclooxygenase)

Citations for Human/Mouse Total COX-2 DuoSet IC ELISA (15)

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Citations are publications that use Bio-Techne products. Selected citations for Human/Mouse Total COX-2 DuoSet IC ELISA include:

Species: Human
Sample Types: Cell Culture Supernates

G Wang et al. (2022), Icariin alleviates uveitis by targeting peroxiredoxin 3 to modulate retinal microglia M1/M2 phenotypic polarization Redox Biology, 52(0):102297.
PMID: 35334248
Species: Human
Sample Types: Cell Culture Supernates

S Yoshida et al. (2022), Green tea catechins inhibit Porphyromonas gulae LPS-induced inflammatory responses in human gingival epithelial cells: Running title Journal of oral biosciences, 0(0).
PMID: 35660639
Species: Mouse
Sample Types: Tissue Homogenates

J Kim et al. (2022), Nilotinib modulates LPS-induced cognitive impairment and neuroinflammatory responses by regulating P38/STAT3 signaling Journal of Neuroinflammation, 19(1):187.
PMID: 35841100
Species: Human
Sample Types: Cell Lysates

B Eggers et al. (2022), Modulation of Inflammatory Responses by a Non-Invasive Physical Plasma Jet during Gingival Wound Healing Cells, 11(17).
PMID: 36078148
Species: Mouse
Sample Types: Tissue Homogenates

JH Liu et al. (2021), Anti-inflammatory effects of Radix aucklandiae herbal preparation ameliorate intestinal mucositis induced by 5-fluorouracil in mice Journal of ethnopharmacology, 271(0):113912.
PMID: 33567307
Species: Human
Sample Types: Cell Lysates

AVB Nogueira et al. (2021), Filifactor alocis and Tumor Necrosis Factor-Alpha Stimulate Synthesis of Visfatin by Human Macrophages International Journal of Molecular Sciences, 22(3).
PMID: 33513808
Species: Human
Sample Types: Cell Culture Lysates

R Nomura et al. (2020), Inhibition of Porphyromonas gulae and periodontal disease in dogs by a combination of clindamycin and interferon alpha Sci Rep, 10(1):3113.
PMID: 32080231
Species: Rat
Sample Types: Tissue Homogenates

CT Hsieh et al. (2018), Systemic Lipopolysaccharide-Induced Pain Sensitivity and Spinal Inflammation Were Reduced by Minocycline in Neonatal Rats Int J Mol Sci, 19(10).
PMID: 30262747
Species: Mouse
Sample Types: Serum

XJ Ma et al. (2018), JIP3 knockout protects mice against high fat diet-induced liver injury Biochem. Biophys. Res. Commun., 0(0).
PMID: 29454969
Species: Rat
Sample Types: Tissue Homogenates

SH Kim et al. (2017), Suppressive Effect of the n-Hexane Extract of Litsea japonica Fruit Flesh on Monosodium-Iodoacetate-Induced Osteoarthritis in Rats Evid Based Complement Alternat Med, 2017(0):1791403.
PMID: 28904551
Species: Human
Sample Types: Cell Lysates

H Wu et al. (2017), Honokiol improved chondrogenesis and suppressed inflammation in human umbilical cord derived mesenchymal stem cells via blocking nuclear factor-?B pathway BMC Cell Biol., 18(1):29.
PMID: 28851291
Species: Mouse
Sample Types: Tissue Homogenates

SR Burks et al. (2017), Anti-inflammatory drugs suppress ultrasound-mediated mesenchymal stromal cell tropism to kidneys Sci Rep, 7(1):8607.
PMID: 28819122
Species: Mouse
Sample Types: Cell Culture Supernates

(2016), Anthocyanins abrogate glutamate-induced AMPK activation, oxidative stress, neuroinflammation, and neurodegeneration in postnatal rat brain J Neuroinflammation, 13(1):286.
PMID: 27821173
Species: Rat
Sample Types: Tissue Homogenates

Jun Lu (2016), Acupuncture ameliorates inflammatory response in a chronic unpredictable stress rat model of depression Brain Res. Bull, 0(0).
PMID: 27915092
Species: Mouse
Sample Types: BALF

Zhang Z et al. (2014), Protective effect of resveratrol against acute lung injury induced by lipopolysaccharide via inhibiting the myd88-dependent Toll-like receptor 4 signaling pathway. Mol Med Rep, 10(1):101-6.
PMID: 24818579

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