Indicador de calcio Calbryte™-520L AM. Calbryte 520L tiene una baja afinidad con los iones de calcio con un Kd ∼91 μM.
Descripción
Indicador Calbryte™-520L, AM
El ensayo de flujo de calcio intracelular es un método ampliamente utilizado para monitorear vías de transducción de señales y detección de alto rendimiento de receptores acoplados a proteínas G (GPCR) y objetivos de canales de calcio.
Luego de la introducción de Fluo-3 en 1989, Fluo-4, Fluo-8 y Cal-520 se desarrollaron posteriormente con relaciones señal-fondo mejoradas y pronto se convirtieron en los indicadores de Ca2+ de elección para microscopía confocal, citometría de flujo y alta aplicaciones de detección de rendimiento. Sin embargo, todavía hay algunas advertencias con Fluo-4. Por ejemplo, al igual que el Fluo-3, el Fluo-4 presenta una retención intracelular deficiente y se requiere el uso de probenecid para evitar que el Fluo-4 cargado en las células se escape de las células.
El uso de probenecid con ensayos de calcio basados en Fluo-4 compromete los resultados del ensayo ya que está bien documentado que probenecid tiene una variedad de efectos celulares complicados. Calbryte 520L, AM es un nuevo indicador de calcio fluorescente y permeable a las células. Al igual que otras cargas de células AM de tinte, el éster AM de Calbryte 520L no es fluorescente y, una vez que ingresa a la célula, es hidrolizado por la esterasa intracelular y se activa.
El indicador activado es una molécula polar que ya no es capaz de difundirse libremente a través de la membrana celular y queda esencialmente atrapada dentro de las células. Calbryte 520L tiene una baja afinidad por los iones de calcio con una Kd ∼ 91 µM. Calbryte 520L produce una señal de fluorescencia brillante en presencia de calcio en altas concentraciones. Tiene la misma longitud de onda de excitación y emisión que Fluo-4; por lo tanto, los mismos ajustes de ensayo de Fluo-4 se pueden aplicar fácilmente a los ensayos de calcio basados en Calbryte 520L.
Nombre en ingles: Calbryte™-520L AM
Importante: Solo para uso en investigación (RUO). Almacenamiento: Congelar a (< -15 °C), Minimizar la exposición a la luz.
| Catalogo | Producto | Presentación |
|---|---|---|
| AAT-20640 | Indicador Calbryte™-520L AM | 10 x 50 ug |
Plataforma
Microscopio de Fluorescencia
| Excitación | FITC |
| Emisión | FITC |
| Placa Recomendada | Pared negra/Fondo claro |
Lector de microplacas de Fluorescencia
| Excitación | 490 |
| Emisión | 525 |
| Cutoff | 515 |
| Placa Recomendada | Pared Negra / fondo claro |
| Especificaciones Instrumento | Modo de lectura inferior / manipulación de liquidos programable |
Espectro
Abrir en Advanced Spectrum Viewer
Propiedades espectrales
| Excitación (nm) | 493 |
| Emisión (nm) | 515 |
| Rendimiento cuántico | 0.751 |
Calculadora
Volumen de DMSO necesario para reconstituir la masa específica de Calbryte™-520L AM a la concentración dada. Tenga en cuenta que el volumen es solo para preparar la solución madre. Consulte el protocolo experimental de muestra para conocer los buffers experimentales/fisiológicos apropiados.
| 0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
| 1 mM | 114.847 µL | 574.237 µL | 1.148 mL | 5.742 mL | 11.485 mL |
| 5 mM | 22.969 µL | 114.847 µL | 229.695 µL | 1.148 mL | 2.297 mL |
| 10 mM | 11.485 µL | 57.424 µL | 114.847 µL | 574.237 µL | 1.148 mL |
Imagen
Figura 1. Ca2+ Dosis-respuesta dependiente de Calbryte 520L
Productos Relacionados
| Name | Excitation (nm) | Emission (nm) | Quantum yield |
| Calbryte™ 520, potassium salt | 493 | 515 | 0.751 |
| Calbryte™ 590, potassium salt | 581 | 593 | – |
| Calbryte™ 630, potassium salt | 607 | 624 | – |
| Calbryte™-520XL, potassium salt | 493 | 515 | 0.751 |
Bibliografía
Calreticulin regulates TGF-β1-induced epithelial mesenchymal transition through modulating Smad signaling and calcium signaling
Authors: Wu, Yanjiao and Xu, Xiaoli and Ma, Lunkun and Yi, Qian and Sun, Weichao and Tang, Liling
Journal: The International Journal of Biochemistry & Cell Biology (2017)
Monosialoganglioside 1 may alleviate neurotoxicity induced by propofol combined with remifentanil in neural stem cells
Authors: Lu, Jiang and Yao, Xue-qin and Luo, Xin and Wang, Yu and Chung, Sookja Kim and Tang, He-xin and Cheung, Chi Wai and Wang, Xian-yu and Meng, Chen and Li, Qing and others, undefined
Journal: Neural Regeneration Research (2017): 945
Obtaining spontaneously beating cardiomyocyte-like cells from adipose-derived stromal vascular fractions cultured on enzyme-crosslinked gelatin hydrogels
Authors: Yang, Gang and Xiao, Zhenghua and Ren, Xiaomei and Long, Haiyan and Ma, Kunlong and Qian, Hong and Guo, Yingqiang
Journal: Scientific Reports (2017): 41781
Dexmedetomidine reduces hypoxia/reoxygenation injury by regulating mitochondrial fission in rat hippocampal neurons
Authors: Liu, Jia and Du, Qing and Zhu, He and Li, Yu and Liu, Maodong and Yu, Shoushui and Wang, Shilei
Journal: Int J Clin Exp Med (2017): 6861–6868
Di (2-ethylhexyl) phthalate-induced apoptosis in rat INS-1 cells is dependent on activation of endoplasmic reticulum stress and suppression of antioxidant protection
Authors: Sun, Xia and Lin, Yi and Huang, Qiansheng and Shi, Junpeng and Qiu, Ling and Kang, Mei and Chen, Yajie and Fang, Chao and Ye, Ting and Dong, Sijun
Journal: Journal of cellular and molecular medicine (2015): 581–594
The effect of mitochondrial calcium uniporter on mitochondrial fission in hippocampus cells ischemia/reperfusion injury
Authors: Zhao, Lantao and Li, Shuhong and Wang, Shilei and Yu, Ning and Liu, Jia
Journal: Biochemical and biophysical research communications (2015): 537–542
Role of mitochondrial calcium uniporter in regulating mitochondrial fission in the cerebral cortexes of living rats
Authors: Liang, Nan and Wang, Peng and Wang, Shilei and Li, Shuhong and Li, Yu and Wang, Jinying and Wang, Min
Journal: Journal of Neural Transmission (2014): 593–600
Propofol and remifentanil at moderate and high concentrations affect proliferation and differentiation of neural stem/progenitor cells
Authors: Li, Qing and Lu, Jiang and Wang, Xianyu and others, undefined
Journal: Neural regeneration research (2014): 2002
Fungus induces the release of IL-8 in human corneal epithelial cells, via Dectin-1-mediated protein kinase C pathways.
Authors: Peng, Xu-Dong and Zhao, Gui-Qiu and Lin, Jing and Jiang, Nan and Xu, Qiang and Zhu, Cheng-Cheng and Qu, Jain-Qiu and Cong, Lin and Li, Hui
Journal: International journal of ophthalmology (2014): 441–447
Increased expression of cell adhesion molecule 1 by mast cells as a cause of enhanced nerve–mast cell interaction in a hapten-induced mouse model of atopic dermatitis
Authors: Hagiyama, M and Inoue, T and Furuno, T and Iino, T and Itami, S and Nakanishi, M and Asada, H and Hosokawa, Y and Ito, A
Journal: British Journal of Dermatology (2013): 771–778
Referencias
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Comparison of human recombinant adenosine A2B receptor function assessed by Fluo-3-AM fluorometry and microphysiometry
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Aplication Notes
A Comparison of Fluorescent Red Calcium Indicators for Detecting Intracellular Calcium Mobilization in CHO Cells
A Meta-Analysis of Common Calcium Indicators
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A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
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FAQ
Are there any calcium indicators that don’t require probenecid (PBC)?
Are there upgraded trypan blue derivatives for cell viability testing?
Can I intracellularly measure mitochondria calcium flux and changes in mitochondria membrane potential at the same time?
Do you offer any products for measuring intracellular calcium concentration or movement by flow cytometry?
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AssayWise
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