Human/Mouse Total COX-2 DuoSet IC ELISA

Catalog # Availability Size / Price Qty
DYC4198-5
DYC4198-2
Human/Mouse Total COX-2 DuoSet IC ELISA Data
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Product Details
Citations (8)
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Human/Mouse Total COX-2 DuoSet IC ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 mins (after plate preparation)
Sample Volume Required
Cell lysates (100 µL)
Sufficient Materials
Kits available for two, 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 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

Data Examples
SPECIFICITY
DYC4198 DuoSet IC ELISA Figure 1
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Figure 1: 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
The amounts of COX-2 as quantified by the Human/Mouse Total COX-2 DuoSet IC ELISA are consistent with the relative amounts of COX-2 determined by qualitative Western blot analysis.
DYC4198 DuoSet IC ELISA Figure 2
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Figure 2: 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.

Data Example

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: 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.

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

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.

8 Citations: Showing 1 - 8
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  1. 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;19(10):.
    Species: Rat
    Sample Types: Tissue Homogenates
  2. 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;0(0):.
    Species: Mouse
    Sample Types: Serum
  3. 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;2017(0):1791403.
    Species: Rat
    Sample Types: Tissue Homogenates
  4. 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;18(1):29.
    Species: Human
    Sample Types: Cell Lysates
  5. 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;7(1):8607.
    Species: Mouse
    Sample Types: Tissue Homogenates
  6. Anthocyanins abrogate glutamate-induced AMPK activation, oxidative stress, neuroinflammation, and neurodegeneration in postnatal rat brain
    J Neuroinflammation, 2016;13(1):286.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. Acupuncture ameliorates inflammatory response in a chronic unpredictable stress rat model of depression
    Authors: Jun Lu
    Brain Res. Bull, 2016;0(0):.
    Species: Rat
    Sample Types: Tissue Homogenates
  8. 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;10(1):101-6.
    Species: Mouse
    Sample Types: BALF

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