Product Introduction

Calcium measurement plays a vital role in various biological research fields. By utilizing fluorescence probes that emit spectral responses upon binding to Ca²⁺, researchers can study intracellular free Ca² concentration changes through instruments like fluorescence microscopes, flow cytometers, and fluorescence microplate readers.

Fluo-3 AM and Fluo-4 AM are the most commonly used visible light-excited calcium indicators in live cell imaging research. However, these two probes have certain limitations: after esterase hydrolysis, they only exhibit moderate fluorescence intensity in living cells. Moreover, achieving optimal cellular calcium response requires stringent cell loading conditions. The emergence of Fluo-20 AM effectively addresses these issues. While maintaining similar spectral wavelengths (Ex/Em = 495/516 nm) and ease of use as Fluo-3 AM, Fluo-4 AM, and Fluo-8 AM, it significantly improves cell loading conditions and calcium response efficiency. For instance, Fluo-20 AM can complete cell loading at room temperature, whereas Fluo-3 AM and Fluo-4 AM require a 37℃°C environment for loading. Additionally, Fluo-20 AM demonstrates four times the brightness of Fluo-4 AM, eight times that of Fluo-3 AM, and twice that of Fluo-8 AM. Fluo-20 AM enables easy detection of weak calcium ion concentration changes while minimizing background interference, making it particularly suitable for scenarios with weak signals such as primary cells or calcium-sensitive cells. As an acetyl methyl ester derivative with cell membrane permeability, it can easily enter cells through simple culture. Once inside the cell, it is cleaved by endolysin into non-permeable Fluo-20, which remains within the cell to perform physiological functions. In summary, Fluo-20 AM serves as an ideal replacement for Fluo-3 AM, Fluo-4 AM, and Fluo-8 AM.

Scope of Application

Calcium ion detection

Product Parameter

Appearance: Solid powder soluble in DMSO

Ex:495 nm

Em:516 nm

Product features

1.Compared to Fluo-3 AM and Fluo-4 AM:

1) Flexible loading conditions: Fluo-8 AM exhibits weaker temperature dependence in loading, maintaining optimal staining performance at both room temperature and 37°C incubation. Unlike the strict 37°C requirement of Fluo-3 AM and Fluo-4 AM, this feature makes it more suitable for high-throughput screening experiments;

2) Exceptional brightness: Fluo-8 AM demonstrates outstanding luminescence, twice as bright as Fluo-4 AM and four times brighter than Fluo-3 AM.

2. Compared to other UV-excited fluorescent probes like Fura-2 and Indo-1:

1) Sample compatibility: It can be excited within the visible light range, avoiding phototoxicity to cells. Many biological samples, especially live cells, are sensitive to UV radiation which may induce photochemical reactions and DNA damage. Visible light excitation better preserves cellular physiological states, making it ideal for prolonged calcium ion monitoring;

2) Superior tissue penetration: UV light has poor tissue penetration in biological systems and is easily absorbed/scattered by tissues. Visible light

Basic Information

Product Components

Note: The staining working solution is calculated as 5 μM per 100 μL used.

Notes

1.The AM ester group is hygroscopic. Always allow it to reach room temperature after removal from the refrigerator before opening. Fluo-20 AM is difficult to observe in trace amounts – perform an immediate centrifugation after reaching room temperature to ensure the powder settles completely at the bottom of the tube.

2. Fluo-20 AM decomposes rapidly upon contact with water. If not used immediately, store in small portions.

3. All fluorescent dyes exhibit quenching effects. Avoid light exposure during experiments to minimize quenching.

4. For safety, wear lab coats and disposable gloves during handling.

5. This product is for scientific research only and must not be used for clinical diagnosis or treatment.

This product is for research use only！