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Txb2 eia kit

Manufactured by Cayman Chemical
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The TXB2 EIA kit is a quantitative enzyme immunoassay designed for the measurement of 11β-Prostaglandin F2α (11β-PGF2α), a metabolite of Thromboxane B2 (TXB2), in biological samples.

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Lab products found in correlation

Adenosine diphosphate (adp), by Merck Group (3 mentions) U46619, by Cayman Chemical (2 mentions) Aspirin, by Merck Group (2 mentions) Meclofenamate, by Merck Group (1 mentions) Microvue complement c3a plus, by Quidel (1 mentions) Synergy ht microplate reader, by Agilent Technologies (1 mentions) Fura 2 am, by Merck Group (1 mentions) Hrp conjugated anti mouse igg, by Cell Signaling Technology (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Heparin, by Merck Group (1 mentions) Thrombin, by Cayman Chemical (1 mentions) Bca protein assay kit, by Thermo Fisher Scientific (1 mentions) U73122, by Cayman Chemical (1 mentions) Sepharose cl 2b, by GE Healthcare (1 mentions) Cgmp eia kit, by Cayman Chemical (1 mentions) Sq29548, by Cayman Chemical (1 mentions) Y 27632, by Cayman Chemical (1 mentions) Chrono lume reagent, by Chrono-Log (1 mentions) Pge2 d4, by Cayman Chemical (1 mentions)

Spelling variants of this product

EIA kit (244 citations) TXB2 enzyme immunoassay (EIA) kit (5 citations)

12 protocols using txb2 eia kit

1

Plasma Thromboxane B2 Quantification

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A competitive enzyme linked immunosorbent assay (EIA) was utilized according to manufacturer's instructions (TXB2 EIA Kit, Cayman Chemical Co., Ann Arbor, MI) in order to analyze thromboxane B2 (TXB2) in citrated plasma. All reagents were prepared fresh using ultra-pure water, and samples diluted as necessary in sample buffer to fall within the standard curve (typically 1:2). No apparent evidence for interference was found in the assay using serially diluted plasma (i.e. <20% difference in the final calculated TXB2 concentration), and therefore did not further purify the plasma samples. The limit of detection (LOD) was 7.8 pg/ml. Stimulated blood was not employed for these analyses as it was the goal of the original sequential therapy trial (Block et al 2013 ), from which this study used data, to determine effects of aspirin and fish oil ingestion alone without additional complication(s) of if and how artificially stimulating them impacted this. Further, a thrombin control was not used since each subject served as their own control in the original analysis (i.e. comparison of TXB2 levels while taking aspirin and/or fish oil compared to TXB2 level while taking neither). All TXB2 assays were analyzed in duplicate, and fell within the standard curve.
Becerra A.Z., Georas S., Brenna J.T., Hopke P.K., Kane C., Chalupa D., Frampton M.W., Block R, & Rich D.Q. (2016). Increases in ambient particulate matter air pollution, acute changes in platelet function, and effect modification by aspirin and omega-3 fatty acids: a panel study. Journal of toxicology and environmental health. Part A, 79(6), 287-298.
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2

Measuring Coagulation and Inflammation Markers

Cited 1 time
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Beta-thromboglobulin (βTG) and prothrombin fragments 1+2 (F1+2) were measured by commercial enzyme-linked immunosorbent assay (ELISA) in plasma samples collected in CTAD tubes as previously described (24 (link)). EDTA plasma was used to measure histamine (ELISA kit Starfish, Gernusco, Italy) and C3a levels. Because the manufacturer of C3a kits recently modified their ELISA to the C3a Plus version (Microvue Complement C3a Plus, Quidel, San Diego, CA), archived GalTKO.hCD46 samples were reanalyzed for this study. Blood, collected at 3 time points (0, 15 and 60min) in EDTA tubes containing 100μl (at 10μg/ml) of meclofenamate (Sigma-Aldrich, St.Louis, MO), was analyzed to measure plasma levels of thromboxane B2 (TXB2 EIA Kit, Cayman Chemical Company, Ann Arbor, MI; Catalog No. 519031).
Burdorf L., Riner A., Rybak E., Salles I., De Meyer S., Shah A., Quinn K., Harris D., Zhang T., Parsell D., Ali F., Schwartz E., Kang E., Cheng X., Sievert E., Zhao Y., Braileanu G., Phelps C., Ayares D., Deckmyn H., Pierson R I.I.I, & Azimzadeh A. (2016). Platelet sequestration and activation during GalTKO.hCD46 pig lung perfusion by human blood is primarily mediated by GPIb, GPIIb/IIIa, and VWF. Xenotransplantation, 23(3), 222-236.
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3

Quantifying TXB2 from Citrated Plasma

Cited 1 time
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A competitive enzyme linked immunosorbent assay (TXB2 EIA Kit, Cayman Chemical Co., Ann Arbor, MI) was used to analyze TXB2 from citrated plasma serially diluted, as necessary, in sample buffer to fall within the standard curve (typically 1:2)[13 (link)].
Block R.C., Abdolahi A., Tu X., Georas S.N., Brenna J.T., Phipps R.P., Lawrence P, & Mousa S.A. (2014). The effects of aspirin on platelet function and lysophosphatidic acids depend on plasma concentrations of EPA and DHA. Prostaglandins, leukotrienes, and essential fatty acids, 96, 17-24.
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4

Measurement of TXB2 Production in Platelets

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Measurement of TXB2 production was conducted using a TXB2 EIA kit (Cayman Chemical Co, Ann Arbor, MI, USA) as previously described (Cho et al., 2011 (link); Seo et al., 2011 (link)). PRP (2×108 platelets/mL) was incubated for 3 min in the presence or absence of samples. Collagen (2 μg/mL) was added and the mixture was incubated at 37°C for 5 min with stirring. EDTA (10 mM) was added to stop TXB2 production. After centrifugation at 12,000 g for 3 min, the amount of TXB2 was measured according to the manufacturer’s instructions.
Kim T.H., Kim H.M., Park S.W, & Jung Y.S. (2015). Inhibitory Effects of Yuzu and Its Components on Human Platelet Aggregation. Biomolecules & Therapeutics, 23(2), 149-155.
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5

Measuring Collagen-Induced Thromboxane B2

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Thromboxane A2 (TXA2) is unstable and instantly converted to thromboxane B2 (TXB2). Therefore, the degree of TXA2 formation in platelets was confirmed by measuring TXB2 using the TXB2 EIA kit (Cayman Chemical Co., Ann Arbor, MI, USA). Rat PRP was pre-incubated at 37 °C for 5 min in the presence of MJGE09 (0, 4, 20, and 100 μg/mL) or aspirin (0.1 mM). Next, collagen (2 μg/mL) was added to PRP and incubated at 37 °C for 5 min, followed by the addition of ethylenediaminetetraacetic acid (EDTA) (10 mM) to stop TXA2 formation. After centrifugation at 12,000× g for 1 min, the supernatant was collected, and the degree of TXB2 formation was measured using the protocol provided in the TXB2 EIA kit.
Jeon Y.D., Lee J.H., Park M.R., Lim J.Y., Kang S.H., Kim D.K, & Lee Y.M. (2021). Gastrodia elata Blume and Zanthoxylum schinifolium Siebold & Zucc Mixed Extract Suppress Platelet Aggregation and Thrombosis. Medicina, 57(10), 1128.
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6

Measurement of Thromboxane B2 Production

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Since thromboxane A2 (TXA2) is unstable and transforms into thromboxane B2 (TXB2) quickly, therefore, TXA2 generation was measured by detecting TXB2 production [7 (link)]. After platelet activation, the reaction was stopped by adding indomethacin (0.2 mM) with EDTA (5 mM). The amounts of TXB2 were determined using a TXB2 EIA kit Cayman Chemical (Ann Arbor, MI, USA). The plate was read at a wavelength of 420 nm using a Synergy HT Microplate Reader (BioTek Instruments, Winoosku, VT., USA).
Shin J.H., Kwon H.W., Irfan M., Rhee M.H, & Lee D.H. (2020). Ginsenoside Rk1 suppresses platelet mediated thrombus formation by downregulation of granule release and αIIbβ3 activation. Journal of Ginseng Research, 45(4), 490-497.
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7

ELISA-based Quantification of Serum and Cell Culture Media

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Serum or cell culture medium was measured by using the TXB2 EIA kit (Cayman Chemical, Cat. #501020) or a mouse VEGF-A ELISA Kit (Pierce, Cat. #EMVEGFA) according to the manufacturer's specifications. Sera were collected from the blood samples of individual mice at the end of the experiment under terminal anesthesia following a protocol for cardiac puncture. Serum samples were separated from blood within 1 h following blood collection by centrifugation at 500g for 10 min, and then aliquotted and stored at −80 °C for subsequent testing. Cell culture media were collected at the end of the experiments and cleared by centrifugation at 17,000g for 10 min and then stored at −80 °C for subsequent testing.
Lichtenberger L.M., Fang D., Bick R.J., Poindexter B.J., Phan T., Bergeron A.L., Pradhan S., Dial E.J, & Vijayan K.V. (2016). Unlocking aspirin’s chemopreventive activity: Role of irreversibly inhibiting platelet cyclooxygenase-1. Cancer prevention research (Philadelphia, Pa.), 10(2), 142-152.
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8

Platelet Activation Pathway Assay Protocol

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U46619, AA, thrombin, SQ29548, Y27632, U73122, SNAP, the cGMP EIA kit and the TxB2 EIA kit were purchased from Cayman Chemical (Ann Arbor, MI, USA). ADP, heparin, PMA, fura 2-AM, bovine serum albumin (BSA) and aspirin were purchased from Sigma (St. Louis, MO, USA). 2,4,6-Trimethyl-N-[3-(trifluoromethyl)phenyl] benzenesulfonamide (m-3M3FBS) was purchased from Calbiochem (Darmstadt, Germany). Antibodies against p44/42 MAPK (ERK1/2), phospho-p44/42 MAPK (pERK1/2) (Thr202/Tyr204), phospho-ROCK2 (Ser1366), and GAPDH, as well as the enhanced chemiluminescence (ECL) system detection kit and HRP-conjugated anti-mouse IgG, were obtained from Cell Signaling Technology (Beverly, MA, USA). The BCA protein assay kit was purchased from Thermo Scientific (Waltham, MA, USA). CHRONO-LUME reagent and collagen were purchased from Chrono-Log Corp (Havertown, PA, USA). The LANCETM cAMP kit was purchased from Perkin Elmer (Waltham, MA, USA). Sepharose CL-2B was purchased from GE Healthcare (Uppsala, Sweden). Compound purification was described in a previous study35 (link). Compounds were dissolved in dimethyl sulfoxide (DMSO). All other chemicals were of reagent grade.
Cong Y., Wang L., Peng R., Zhao Y., Bai F., Yang C., Liu X., Wang D., Ma B, & Cong Y. (2016). Timosaponin AIII induces antiplatelet and antithrombotic activity via Gq-mediated signaling by the thromboxane A2 receptor. Scientific Reports, 6, 38757.
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9

Platelet Aggregation and TXB2 Measurement

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Washed platelets were pre-incubated with experimental samples and stimulated for aggregation reaction as previously described in this paper. The reactions were terminated by adding ice-cold 2.5 mM EGTA and 100 μM indomethacin. After centrifugation at 12000 x g for 3 min at 4 °C, supernatants were collected and TXB2 concentration was measured with TXB2 EIA kit (Cayman, USA).
Son E., Kim S.H., Yang W.K., Kim D.S, & Cha J. (2017). Antiplatelet mechanism of an herbal mixture prepared from the extracts of Phyllostachys pubescens leaves and Prunus mume fruits. BMC Complementary and Alternative Medicine, 17, 541.
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10

Licochalcones Inhibit Prostaglandin Synthesis

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Licochalcones were provided by Research Laboratory of Minophagen Pharmaceutical Co. Ltd. (Zama, Japan). Prostaglandin (PG) E2, d4-PGE2, U46619, COX-1, COX-2 and TXB2 EIA Kit were purchased from Cayman Chemical Company (Ann Arbor, MI). Collagen (Collagenreagent Horm) was purchased from Nycomed Pharma GMBH (Marburg, Germany). Thrombin was purchased from Wako Pure Chemicals (Osaka, Japan). Arachidonic acid and ADP were purchased from Sigma-Aldrich (St. Louis, MO) or Cayman Chemical Company. All other chemicals used were of reagent grade or the highest quality available.
Okuda-Tanino A., Sugawara D., Tashiro T., Iwashita M., Obara Y., Moriya T., Tsushima C., Saigusa D., Tomioka Y., Ishii K, & Nakahata N. (2017). Licochalcones extracted from Glycyrrhiza inflata inhibit platelet aggregation accompanied by inhibition of COX-1 activity. PLoS ONE, 12(3), e0173628.
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