{"id":3244,"date":"2023-01-12T10:49:13","date_gmt":"2023-01-12T16:49:13","guid":{"rendered":"https:\/\/nuevo22.cidsamexico.com\/?post_type=al_product&p=3244"},"modified":"2023-01-20T13:20:01","modified_gmt":"2023-01-20T19:20:01","slug":"screen-quest-fluo-8-medium-removal-calcium-assay-kit-optimized-for-difficult-cell-lines","status":"publish","type":"al_product","link":"https:\/\/nuevo22.cidsamexico.com\/index.php\/productos\/screen-quest-fluo-8-medium-removal-calcium-assay-kit-optimized-for-difficult-cell-lines\/","title":{"rendered":"Screen Quest\u2122 Fluo-8 Medium Removal Calcium Assay Kit *Optimized for Difficult Cell Lines*"},"content":{"rendered":"\n

Los ensayos de flujo de calcio son m\u00e9todos preferidos en el descubrimiento de f\u00e1rmacos para la detecci\u00f3n de receptores acoplados a prote\u00edna G (GPCR). El kit de ensayo de calcio Screen Quest\u2122 Fluo-8 NW proporciona un ensayo homog\u00e9neo basado en fluorescencia para detectar la movilizaci\u00f3n de calcio intracelular. Las c\u00e9lulas que expresan un GPCR de inter\u00e9s que emite se\u00f1ales a trav\u00e9s del calcio est\u00e1n precargadas con nuestro Fluo-8 NW patentado que puede atravesar la membrana celular. <\/p>\n\n\n\n

Fluo-8 NW es el indicador de calcio m\u00e1s brillante disponible para la detecci\u00f3n de HTS. Una vez dentro de la c\u00e9lula, los grupos bloqueadores lipof\u00edlicos de Fluo-8 NW son escindidos por la esterasa celular no espec\u00edfica, lo que da como resultado un tinte fluorescente con carga negativa que permanece dentro de las c\u00e9lulas y su fluorescencia aumenta considerablemente al unirse al calcio. Cuando las c\u00e9lulas se estimulan con compuestos de detecci\u00f3n, el receptor se\u00f1ala la liberaci\u00f3n de calcio intracelular, lo que aumenta en gran medida la fluorescencia de Fluo-8 NW. Las caracter\u00edsticas de su longitud de onda larga, alta sensibilidad y aumento de fluorescencia de 100 a 250 veces (cuando forma complejos con calcio) hacen de Fluo-8 NW un indicador ideal para la medici\u00f3n de calcio celular. <\/p>\n\n\n\n

Este kit de ensayo de calcio Screen Quest Fluo-8 NW proporciona un m\u00e9todo de ensayo optimizado para monitorear los receptores acoplados a prote\u00edna G (GPCR) y los canales de calcio. El ensayo se puede realizar en un conveniente formato de placa de microtitulaci\u00f3n de 96 pozos o de 384 pozos y se puede adaptar f\u00e1cilmente a la automatizaci\u00f3n.  Se vende en Kit de 1, 10 y 100 placas.<\/p>\n\n\n\n

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Catalogo<\/th>Producto<\/th>Presentaci\u00f3n<\/th><\/tr><\/thead>
AAT-36307<\/td>Screen Quest\u2122 Fluo-8 Medium Removal Calcium Assay Kit *Optimized for Difficult Cell Lines*<\/td>1 placa<\/td><\/tr>
AAT-36308<\/td>Screen Quest\u2122 Fluo-8 Medium Removal Calcium Assay Kit *Optimized for Difficult Cell Lines*<\/td>10 placas<\/td><\/tr>
AAT-36309<\/td>Screen Quest\u2122 Fluo-8 Medium Removal Calcium Assay Kit *Optimized for Difficult Cell Lines*<\/td>100 placas<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
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\"pdf\"\/<\/figure>\n\n\n\n

SDS<\/a><\/p>\n\n\n\n

\"pdf\"\/<\/figure>\n\n\n\n

Protocol<\/a><\/p>\n<\/div>\n\n\n\n

Importante: Solo para uso en investigaci\u00f3n (RUO). <\/p>\n\n\n\n

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Plataforma<\/mark><\/p>\n\n\n\n

Lector de Microplacas de Fluorescencia<\/mark> <\/p>\n\n\n\n

Excitaci\u00f3n<\/td>490 nm<\/td><\/tr>
Emisi\u00f3n<\/td>525 nm<\/td><\/tr>
Cutoff<\/td>510 nm<\/td><\/tr>
Placa recomendada<\/td>Pared negra \/ fondo claro <\/td><\/tr>
Especificaciones instrumento<\/td>Modo de lectura inferior\/Manejo de l\u00edquidos programable<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
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Componentes<\/mark><\/p>\n\n\n\n

Componente A: Fluo-8 NW<\/td>10 viales liofilizados<\/td><\/tr>
Componente B: 10X Pluronic\u00ae F127 Plus<\/td>10 botellas (10 mL\/botella)<\/td><\/tr>
Componente C: HHBS (Hanks’ buffer con 20 mM Hepes)<\/td>No Incluido<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n
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Otros Instrumentos<\/mark><\/p>\n\n\n\n

ArrayScan, FDSS, FLIPR, FlexStation, IN Cell Analyzer, NOVOStar, ViewLux<\/p>\n\n\n\n

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Espectro<\/mark> <\/p>\n\n\n\n

Abrir en Advanced Spectrum Viewer<\/a><\/em><\/p>\n\n\n\n

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Propiedades espectrales<\/mark><\/p>\n\n\n\n

Factor de correcci\u00f3n (260 nm)<\/td>1.076<\/td><\/tr>
Factor de correcci\u00f3n (280 nm)<\/td>0.769<\/td><\/tr>
Coeficiente de Extinci\u00f3n (cm -1<\/sup> M -1<\/sup>)<\/td>23430<\/td><\/tr>
Excitaci\u00f3n (nm)<\/td>495<\/td><\/tr>
Emisi\u00f3n (nm)<\/td>516<\/td><\/tr>
Rendimiento cu\u00e1ntico <\/td>0.161<\/sup><\/td><\/tr><\/tbody><\/table>
1<\/sup> Calcium bound (pH 7.2)<\/figcaption><\/figure>\n\n\n\n
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PREPARACION DE SOLUCION DE STOCK<\/p>\n\n\n\n

A menos que se indique lo contrario, todas las soluciones madre no utilizadas deben dividirse en al\u00edcuotas de un solo uso y almacenarse a -20 \u00b0C despu\u00e9s de la preparaci\u00f3n. Evite los ciclos repetidos de congelaci\u00f3n y descongelaci\u00f3n.<\/em><\/p>\n\n\n\n

1. Soluci\u00f3n madre Fluo-8 NW:<\/em>
Para Cat No. 36307, agregue 10 \u00b5l de DMSO en Fluo-8 NW (componente A) y m\u00e9zclelos bien.
Para Cat No. 36308 y 36309, agregue 100 \u00b5L de DMSO en Fluo-8 NW (Componente A) y mezcle bien. Nota: 10 \u03bcL de soluci\u00f3n madre Fluo-8 NW es suficiente para 1 placa.<\/p>\n\n\n\n

2. Soluci\u00f3n madre de buffer de ensayo (1X):<\/em>
Para los n\u00fameros de cat\u00e1logo 36307 y 36308, agregue 9 ml de HHBS (componente C) en 10X Pluronic\u00ae F127 Plus (1 ml, componente B) y mezcle bien.
Para Cat No. 36309, agregue el frasco completo de 10X Pluronic\u00ae F127 Plus (10 ml, componente B) en 90 ml de buufer HHBS (no incluido en el kit) y mezcle bien. Nota: 10 ml de buffer de ensayo 1X son suficientes para una placa.<\/p>\n\n\n\n

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PREPARACION DE SOLUCION DE TRABAJO<\/p>\n\n\n\n

Agregue 10 \u00b5l de soluci\u00f3n madre de Fluo-8 NW DMSO a 10 ml de Buffer de ensayo 1X y mezcle bien. Esta soluci\u00f3n de trabajo es estable durante al menos 2 horas a temperatura ambiente.<\/p>\n\n\n\n

Para guia sobre la preparaci\u00f3n de muestras de c\u00e9lulas, visite:<\/p>\n\n\n\n

https:\/\/www.aatbio.com\/resources\/guides\/cell-sample-preparation.html<\/a><\/p>\n\n\n\n

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<\/p>\n<\/div><\/div>\n\n\n\n

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Imagen<\/mark><\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

Figura 1<\/strong>. La respuesta a la dosis de carbacol se midi\u00f3 en c\u00e9lulas HEK-293 con el kit de ensayo Screen Quest\u2122 Fluo-8 NW y el kit de ensayo Fluo-4 NW. Se sembraron c\u00e9lulas HEK-293 40.000 c\u00e9lulas\/100 \u00b5l\/pozo, durante la noche a en una placa de 96 pozos Costar de pared negra\/fondo transparente. Se elimin\u00f3 el medio de crecimiento y las c\u00e9lulas se incubaron con 100 \u00b5l de soluci\u00f3n de carga de colorante utilizando el kit de ensayo de calcio Screen Quest\u2122 Fluo 8-NW o el kit Fluo-4 NW (seg\u00fan las instrucciones del fabricante) durante 1 hora a temperatura ambiente. NOVOstar (BMG Labtech) a\u00f1adi\u00f3 carbacol (25 \u00b5l\/pozo) para alcanzar las concentraciones finales indicadas. La EC50 de Carbacol usando Fluo8 NW es de aproximadamente 1,2 \u00b5M.<\/p>\n\n\n\n

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Bibliograf\u00eda<\/mark><\/p>\n\n\n\n

Ver todas las\u00a0158 bibliograf\u00edas:<\/mark>\u00a0<\/em>Citation Explorer<\/a><\/p>\n\n\n\n

Renal tubular epithelial TRPA1 acts as an oxidative stress sensor to mediate ischemia-reperfusion-induced kidney injury through MAPKs\/NF-$\\kappa$B Signaling<\/a>
Authors: <\/strong>Wu, Chung-Kuan and Wu, Chia-Lin and Lee, Tzong-Shyuan and Kou, Yu Ru and Tarng, Der-Cherng
Journal: <\/strong>International journal of molecular sciences (2021): 2309<\/p>\n\n\n\n

Cellular repair mechanisms triggered by exposure to silver nanoparticles and ionic silver in embryonic zebrafish cells<\/a>
Authors: <\/strong>Quevedo, Ana C and Lynch, Iseult and Valsami-Jones, Eugenia
Journal: <\/strong>Environmental Science: Nano (2021)<\/p>\n\n\n\n

Lung epithelial TRPA1 mediates lipopolysaccharide-induced lung inflammation in bronchial epithelial cells and mice<\/a>
Authors: <\/strong>Ko, Hsin-Kuo and Lin, An-Hsuan and Perng, Diahn-Warng and Lee, Tzong-Shyuan and Kou, Yu Ru
Journal: <\/strong>Frontiers in physiology (2020): 596314<\/p>\n\n\n\n

MMP-12 activates protease-activated receptor-1, upregulates placenta growth factor, and leads to pulmonary emphysema
Authors: <\/strong>Hou, Hsin-Han and Wang, Hao-Chien and Cheng, Shih-Lung and Chen, Yen-Fu and Lu, Kai-Zen and Yu, Chong-Jen
Journal: <\/strong>American Journal of Physiology-Lung Cellular and Molecular Physiology (2018): L432–L442<\/p>\n\n\n\n

Human lysophosphatidic acid receptor 2-K1. 31 and K7. 36 gatekeeper functions provide new insight into robust affinity for LPA-type agonists.<\/a>
Authors: <\/strong>Omotuyi, Olaposi I
Journal: <\/strong>Journal of Systems Biology & Proteome Research (2017)<\/p>\n\n\n\n

Cells smell on a CMOS: A portable odorant detection system using cell-laden collagen pillars<\/a>
Authors: <\/strong>Hirata, Yusuke and Morimoto, Yuya and Nam, Eunryel and Yoshida, Shotaro and Takeuchi, Shoji
Journal: <\/strong>(2017): 13–16<\/p>\n\n\n\n

Z-360 Suppresses Tumor Growth in MIA PaCa-2-bearing Mice via Inhibition of Gastrin-induced Anti-Apoptotic Effects<\/a>
Authors: <\/strong>SHIOMI, YOSHIHIRO and YOSHIMURA, MAKOTO and KUKI, KAZUMASA and HORI, YUKO and TANAKA, TAKAO
Journal: <\/strong>Anticancer Research (2017): 4127–4137<\/p>\n\n\n\n

Laminarin counteracts diet-induced obesity associated with glucagon-like peptide-1 secretion<\/a>
Authors: <\/strong>Yang, Liusong and Wang, Lina and Zhu, Canjun and Wu, Junguo and Yuan, Yexian and Yu, Lulu and Xu, Yaqiong and Xu, Jingren and Wang, Tao and Liao, Zhengrui and others, undefined
Journal: <\/strong>Oncotarget (2017): 99470<\/p>\n\n\n\n

2-OMe-lysophosphatidylcholine analogues are GPR119 ligands and activate insulin secretion from \u03b2TC-3 pancreatic cells: Evaluation of structure-dependent biological activity<\/a>
Authors: <\/strong>Drzazga, Anna and Sowi\u0144ska, Agata and Krzemi\u0144ska, Agnieszka and Okruszek, Andrzej and Paneth, Piotr and Koziolkiewicz, Maria and Gendaszewska-Darmach, Edyta
Journal: <\/strong>Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids (2017)<\/p>\n\n\n\n

L-Type Calcium Channel Inhibition Contributes to the Proarrhythmic Effects of Aconitine in Human Cardiomyocytes<\/a>
Authors: <\/strong>Wu, Jianjun and Wang, Xiangchong and Chung, Ying Ying and Koh, Cai Hong and Liu, Zhenfeng and Guo, Huicai and Yuan, Qiang and Wang, Chuan and Su, Suwen and Wei, Heming
Journal: <\/strong>PloS one (2017): e0168435<\/p>\n\n\n\n

<\/p>\n\n\n\n

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Referencias<\/mark><\/p>\n\n\n\n

Ver todas las 26 referencias:\u00a0<\/em>Citation Explorer<\/a><\/p>\n\n\n\n

Novel fluo-4 analogs for fluorescent calcium measurements
Authors: <\/strong>Martin VV, Beierlein M, Morgan JL, Rothe A, Gee KR.
Journal: <\/strong>Cell Calcium (2004): 509<\/p>\n\n\n\n

Kinetic characterization of novel NR2B antagonists using fluorescence detection of calcium flux
Authors: <\/strong>Bednar B, Cunningham ME, Kiss L, Cheng G, McCauley JA, Liverton NJ, Koblan KS.
Journal: <\/strong>J Neurosci Methods (2004): 247<\/p>\n\n\n\n

Flow cytometric kinetic assay of calcium mobilization in whole blood platelets using Fluo-3 and CD41
Authors: <\/strong>do Ceu Monteiro M, Sansonetty F, Goncalves MJ, O’Connor JE.
Journal: <\/strong>Cytometry (1999): 302<\/p>\n\n\n\n

Amplitude distribution of calcium sparks in confocal images: theory and studies with an automatic detection method
Authors: <\/strong>Cheng H, Song LS, Shirokova N, Gonzalez A, Lakatta EG, Rios E, Stern MD.
Journal: <\/strong>Biophys J (1999): 606<\/p>\n\n\n\n

A simple numerical model of calcium spark formation and detection in cardiac myocytes
Authors: <\/strong>Smith GD, Keizer JE, Stern MD, Lederer WJ, Cheng H.
Journal: <\/strong>Biophys J (1998): 15<\/p>\n\n\n\n

Monitoring calcium in outer hair cells with confocal microscopy and fluorescence ratios of fluo-3 and fura-red
Authors: <\/strong>Su ZL, Li N, Sun YR, Yang J, Wang IM, Jiang SC.
Journal: <\/strong>Shi Yan Sheng Wu Xue Bao (1998): 323<\/p>\n\n\n\n

Loading and localization of Fluo-3 and Fluo-3\/AM calcium indicators in sinapis alba root tissue
Authors: <\/strong>Tretyn A, Kado RT, Kendrick RE.
Journal: <\/strong>Folia Histochem Cytobiol (1997): 41<\/p>\n\n\n\n

Nucleoplasmic and cytoplasmic differences in the fluorescence properties of the calcium indicator Fluo-3
Authors: <\/strong>Perez-Terzic C, Stehno-Bittel L, Clapham DE.
Journal: <\/strong>Cell Calcium (1997): 275<\/p>\n\n\n\n

Detection of a trigger zone of bradykinin-induced fast calcium waves in PC12 neurites
Authors: <\/strong>Reber BF, Schindelholz B.
Journal: <\/strong>Pflugers Arch (1996): 893<\/p>\n\n\n\n

Improved four-color flow cytometry method using fluo-3 and triple immunofluorescence for analysis of intracellular calcium ion ([Ca2+]i) fluxes among mouse lymph node B- and T-lymphocyte subsets
Authors: <\/strong>Greimers R, Trebak M, Moutschen M, Jacobs N, Boniver J.
Journal: <\/strong>Cytometry (1996): 205<\/p>\n\n\n\n

<\/div>\n\n\n\n

Application Notes (en Ingles)<\/mark><\/p>\n\n\n\n

A Comparison of Fluorescent Red Calcium Indicators for Detecting Intracellular Calcium Mobilization in CHO Cells<\/a>
A Meta-Analysis of Common Calcium Indicators<\/a>
A New Red Fluorescent & Robust Screen Quest\u2122 Rhod-4\u2122 Ca2+Indicator for Screening GPCR & Ca2+ Channel Targets<\/a>
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets<\/a>
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets<\/a><\/p>\n\n\n\n

<\/div>\n","protected":false},"excerpt":{"rendered":"

Kit de ensayo de medio para eliminacion de calcio Screen Quest\u2122 Fluo-8. Optimizado para lineas celulares dificiles. Este kit de ensayo de calcio Screen Quest\u2122 Fluo-8 NW proporciona un ensayo homog\u00e9neo basado en fluorescencia para detectar la movilizaci\u00f3n de calcio intracelular. <\/p>\n","protected":false},"featured_media":3246,"template":"","al_product-cat":[34],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product\/3244"}],"collection":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product"}],"about":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/types\/al_product"}],"version-history":[{"count":4,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product\/3244\/revisions"}],"predecessor-version":[{"id":3721,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product\/3244\/revisions\/3721"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/media\/3246"}],"wp:attachment":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/media?parent=3244"}],"wp:term":[{"taxonomy":"al_product-cat","embeddable":true,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/al_product-cat?post=3244"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}