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Pge2 elisa

Manufactured by Cayman Chemical
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The PGE2 ELISA is a quantitative enzyme-linked immunosorbent assay kit designed to measure the concentration of Prostaglandin E2 (PGE2) in various sample types. The kit utilizes a competitive binding technique and includes all the necessary reagents and materials to perform the assay.

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Lab products found in correlation

Arachidonic acid, by Merck Group (2 mentions) Rneasy mini kit, by Qiagen (1 mentions) Qiashredder, by Qiagen (1 mentions) Beacon designer software 7, by Premier Biosoft (1 mentions) Human ldl, by Lee Biosolutions (1 mentions) Cay10598, by Cayman Chemical (1 mentions) Collagenase type 4, by Merck Group (1 mentions) Hyaluronidase type 5, by Merck Group (1 mentions) Celecoxib, by Abcam (1 mentions) β actin antibody, by Merck Group (1 mentions) Cox 2 antibody, by BD (1 mentions) Dnase type 4, by Merck Group (1 mentions) Infinite m2000, by Tecan (1 mentions)

Spelling variants of this product

PGE2 ELISA kit (65 citations) PGE2 ELISA kit Monoclonal (9 citations) Prostaglandin E2 (PGE2) ELISA kit (8 citations) PGE2 Express ELISA Kit (7 citations) PGE2 enzyme-linked immunosorbent assay (ELISA) kit (4 citations) ELISAs (3 citations) ELISA kits for PGE2 (3 citations) PGE2-specific ELISA kit (3 citations) PGE2 ELISA assay (2 citations)

6 protocols using pge2 elisa

1

Quantification of Inflammatory Mediators

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NO concentration in the cell media was determined using the Griess reagent according to the manufacturer’s protocol and using NaNO2 standard curve. Prostaglandin E2 (PGE2), tumor necrosis alpha (TNF-α), and monocyte chemoattractant protein 1 (MCP-1) were quantified in the cell culture media using commercially available kits following the manufacturer’s instructions (PGE2 ELISA from Cayman Chemical (Ann Arbor, MI, USA) and TNF-α and MCP-1 ELISA kits from R&D systems (Minneapolis, MN, USA)).
Rebollo-Hernanz M., Zhang Q., Aguilera Y., Martín-Cabrejas M.A, & Gonzalez de Mejia E. (2019). Relationship of the Phytochemicals from Coffee and Cocoa By-Products with their Potential to Modulate Biomarkers of Metabolic Syndrome In Vitro. Antioxidants, 8(8), 279.
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2

siRNA-Mediated COX-2 Silencing and PGE2 Regulation

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The siRNA/compound 4 nanoplex in RPMI 1640 medium solution (concentration of siRNA: 100 nM, N/P = 15) was added to each dish for 8 h incubation. After incubation, cells were incubated in fresh medium for a further 16 h. In TPA treatment experiments, TPA was added to medium at 6 h before harvest. Before harvesting the cells, the supernatant of cell culture medium was collected, and PGE2 ELISA was performed following instructions provided by the manufacturer (Cayman Chemical, Ann Arbor, MI). ELISA was performed in triplicates. Total RNA was isolated from cells by using QIA shredder and RNeasy Mini kit (Qiagen, Valencia, CA, USA) according to the manufacturer's protocol. The expression of target RNA relative to the housekeeping gene hypoxanthine phosphoribosyl transferase 1 (HPRT1) was calculated based on the threshold cycle (Ct) as R = 2 - Δ(ΔCt), where ΔCt = Ct of target - Ct of HPRT1. Primer for COX-2 was purchased from Qiagen (Cat. No. QT00040586). The following primers against HPRT1-the house keeping gene-Fwd-5'-CCTGGCGTCGTGATTAGTGATG-3' and Rev-5'-CAGAGGGCTACAATGTGATGGC-3' were designed using either Beacon designer software 7.8 (Premier Biosoft, Palo Alto, CA, USA) or a free web-based software Primer3Plus.
Chen Z., Krishnamachary B., Penet M.F, & Bhujwalla Z.M. (2018). Acid-degradable Dextran as an Image Guided siRNA Carrier for COX-2 Downregulation. Theranostics, 8(1), 1-12.
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3

Prostaglandin E2 Signaling Assay

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All chemicals were purchased from commercial sources indicated throughout the text. Oligonucleotides were custom-synthesized by Biolegio (Nijmegen, Netherlands).
EP receptor specific agonists 19(R)-hydroxy prostaglandin E2 (EP2 agonist), CAY10598 (EP4 agonist), and the PGE2 ELISA were from Cayman chemicals (Ann Arbor, USA). Antibodies used were GAPDH (sc-2578) and mPGES-1 (sc-365844) from Santa Cruz Biotechnology (Heidelberg, Germany), COX-2 (12282), Akt (9272) and pAkt Ser-473 (4060), ERK (9102), pERK (9106), pIKK α/β Ser176/180 (2694), and cleaved caspase-3 Asp175 (9661) from Cell Signalling (Frankfurt, Germany). Human LDL was purchased from LEE Biosolutions (Maryland Heights, USA).
Neuschäfer-Rube F., Schön T., Kahnt I, & Püschel G.P. (2023). LDL-Dependent Regulation of TNFα/PGE2 Induced COX-2/mPGES-1 Expression in Human Macrophage Cell Lines. Inflammation, 46(3), 893-911.
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4

Inflammatory Mediators in Tumor Microenvironment

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Tumor homogenates were harvested from mice treated as indicated, and mechanically disaggregated and digested with triple enzyme mixture (Collagenase type IV, DNase type IV, and Hyaluronidase type V (Sigma-Aldrich, St Louis, MO)). Serum was collected through sub-mandibular bleed. In vitro cells infected with virus at MOI=1 for 24 hr or pretreated for 24 hr with 20 nm Celecoxib (BioVision, San Francisco, CA) were pretreated with 20 μm arachidonic acid (Sigma-Aldrich, St. Louis, MO) 4 hr prior to harvest. All experiments were performed in triplicate. Mouse SDF-1(CXCL12), RANTES (CCL5) and I-TAC(CXCL11) ELISA were performed according to the manufacturer’s protocol (Abcam, Cambridge, MA) and PGE2 ELISA were performed according to the manufacturer’s protocol (Cayman, Ann Arbor, Michigan) and optical density was detected with the 3,3,5,5-tetramethylbenzidine (TMB) peroxidase substrate kit (Vector Labs, Burlingame, CA).
For western blot assay, in vitro lysed cell protein was prepared. Mouse HPGD antibody (Abcam,Cambridge, MA), COX2 antibody (BD, Franklin Lakes, New Jersey) and beta-actin antibody (Sigma-Aldrich, St. Louis, MO) were used for Western Blot according to the manufacturer’s protocol.
Hou W., Sampath P., Rojas J.J, & Thorne S.H. (2016). Oncolytic virus-mediated targeting of PGE2 in the tumor alters the immune status and sensitizes established and resistant tumors to immunotherapy. Cancer cell, 30(1), 108-119.
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5

PGE2 Measurement Protocol

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For PGE2 measurements, cells were first stimulated with sEV for 24 h and then treated with 10 μM arachidonic acid (AA, Sigma‐Aldrich, Darmstadt, GER). 15 min after the addition of AA, supernatants were harvested and centrifuged at 17,000×g and 4°C for 5 min to eliminate cell debris. Supernatants were transferred to fresh tubes and PGE2 concentrations were measured using a PGE2 ELISA (514010, Cayman Chemicals, Ann Arbor, USA) according to the manufacturer's instructions. All samples were measured in duplicates. The absorbance was determined with the Tecan Infinite M 2000 (Tecan Group, Männedorf, CH).
Donzelli J., Proestler E., Riedel A., Nevermann S., Hertel B., Guenther A., Gattenlöhner S., Savai R., Larsson K, & Saul M.J. (2021). Small extracellular vesicle‐derived miR‐574‐5p regulates PGE2‐biosynthesis via TLR7/8 in lung cancer. Journal of Extracellular Vesicles, 10(12), 12143.
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6

Quantifying PGE2 levels in cell cultures

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The determination of prostaglandins E2 was performed by the competitive enzyme-linked immunosorbent assay (ELISA) on culture supernatants after pretreatment and subsequent activation of the cells with LPS/IFN-γ using the commercial Cayman PGE-2 ELISA KIT Monoclonal.
Boukhers I., Boudard F., Morel S., Servent A., Portet K., Guzman C., Vitou M., Kongolo J., Michel A, & Poucheret P. (2022). Nutrition, Healthcare Benefits and Phytochemical Properties of Cassava (Manihot esculenta) Leaves Sourced from Three Countries (Reunion, Guinea, and Costa Rica). Foods, 11(14), 2027.
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