RatioWorks™ BCFL Acid *Superior replacement for BCECF*

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Unit Size: Cat No: Price (USD): Qty:
1 mg 21189 $95


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Additional Ordering Information
Telephone: 1-800-990-8053
Fax: 1-408-733-1304
Email: sales@aatbio.com
International: See distributors





Overview

Ex/Em (nm)503/528
MW~500
SolventDMSO
Storage F/D/L
Category Cell Biology
pH and Ion Indicators
Related Cell Metabolism
Secondary Reagents
Intracellular pH plays an important modulating role in many cellular events, including cell growth, calcium regulation, enzymatic activity, receptor-mediated signal transduction, ion transport, endocytosis, chemotaxis, cell adhesion and other cellular processes. pH-sensitive fluorescent dyes have been widely applied to monitor changes in intracellular pH in recent years. Imaging techniques that use fluorescent pH indicators also allow researchers to investigate these processes with much greater spatial resolution and sampling density that can be achieved using other technologies such as microelectrode. Among them, 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) is the most popular pH probe since it can be used to monitor cellular pH ratiometrically. However, all the commercial BCECF AM is a complex mixture of at least three isomers with different ratios from batch to batch, complicating the BCECF applications. BCFL is developed to overcome this isomer difficulty associated with BCECF AM. As BCECF, BCFL exhibits a pH-dependent dual excitation, essentially identical to BCECF. It has pKa of ~7.0, identical to BCECF too. As with BCECF, the dual excitation spectrum of BCFL with an isosbestic point at 454 nm should make BCFL a good excitation-ratiometrie pH indicator. BCFL ratiometric imaging makes intracellular pH determination essentially independent of several variable factors, including dye concentration, path length, cellular leakage and photobleaching rate. BCFL, AM is a single isomer, making the pH measurement much more reproducible than the BCECF, AM, which is consisted of quite a few different isomers. This BCFL is the esterase-hydrolyzed product of BCFL AM that can be used a reference standard with BCFL AM.




Calculators
Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of RatioWorks™ BCFL Acid *Superior replacement for BCECF* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.



Molarity calculator

Table 2. Enter any two values (mass, volume, concentration) to calculate the third.

Mass Molecular weight Volume Concentration Moles
/ = x =
 










Protocol


Quick Preview

This protocol only provides a guideline, and should be modified according to your specific needs.

1.       Make a series 50 mM phosphate buffer from pH 3 to 10 (3, 4, 5, 6, 7, 7.5, 8, 10)

2.       Make 1 mM DMSO stock solution. The DMSO stock solution is good for 6 months if stored at -20 °C.

3.       Make 1: 1000 dilution  into different pH buffer to have 1uM.

4.       Measure the fluorescent ratio at Ex/Em = 500/535 and 430/535 nm .






References & Citations

The F0F1 ATP synthase regulates human neutrophil migration through cytoplasmic proton extrusion coupled with ATP generation
Authors: Jun Gao, Tian Zhang, Zhanfang Kang, Weijen Ting, Lingqing Xu, Dazhong Yin
Journal: Molecular Immunology (2017): 219--226

Oxidative Stress-Activated NHE1 Is Involved in High Glucose-Induced Apoptosis in Renal Tubular Epithelial Cells
Authors: Yiqing Wu, Min Zhang, Rui Liu, Chunjie Zhao
Journal: Yonsei Medical Journal (2016): 1252--1259






Additional Documents

 
Safety Data Sheet (SDS)


Catalogs
1. Physiological Probes & Assay Kits

Application Notes
1. AssayWise Letters 2013, Vol 2(2)

Certificate of Analysis