- Description
- Additional Information
- Readable Documents
- Assay Principle
- Reviews
Introduction
The two new novel probes, Aminophenyl fluorescein (APF) and Hydroxyphenyl fluorescein (HPF) developed by Tetsuo Nagano et. al. (1), are selective for the detection of highly reactive oxygen species (hROS). Both probes have little reactivity towards other ROS such as: singlet oxygen (021), superoxide (O2-•), hydrogen peroxide (H202), nitric oxide (NO•), and alkyl peroxide (RO2•) (see table below)1. HPF/APF are cell permeable and can be used in combination to detect hypochlorite (-OCl) production in cells (see fig 1). Hypochlorite can be detected by loading two samples, one with APF and the other with HPF. Hypochlorite production is visualized by increase in fluorescence of APF loaded cells and no increase in fluorescence in HPF loaded cells.
Key Benefits
- Quenched Cell permeable dye.
- One Step, No wash Homogenous assay.
- Adaptable to High throughput assay platforms.
- Can monitor multiple time points to follow real time kinetics.
- Non-destructive cell based assay allows monitoring of additional parameters.
- Applications – Fluorescent plate reader / Flow Cytometery / Fluorescent Microscopy.
Additional information
| Kit Size | 100 |
|---|



Figure : Detection of Hypochlorite ( -OCl ) in neutrophils. Neutrophils were isolated from porcine blood, washed in Krebs-Ringers phosphate buffer (as described in step V 1. above) and seeded in glass bottom dishes. The cells were then loaded with APF or HPF (10mM final) by incubation for 30 minutes at room temperature. The Dye-loaded neutrophils were stimulated with PMA (2ng/mL). Fluorescence images were acquired before and 10 minutes after stimulation. Excitation: 488nm emission: 505-550 nm barrier filters1. Hypochlorite production can be detected with increase APF fluorescence and no increase in HPF fluorescence.
| Document Title |
| OclProtocol |
| OCI Datasheet |
| msds.APF-HPF-OCl |
| Reference |
| Ken-ichi Setsukinai, Yasuteru Urano, Katsuko Kakinuma, Hideyuki J. Majima , and Tetsuo Nagano. Development of Novel Fluorescence Probes That Can Reliably Detect Reactive Oxygen Species and Distinguish Specific Species. THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 278, No. 5, Issue of January 31, pp. 3170–3175, 2003 |
| Part# | Reagent | Temperature |
| Part# 4012 | HPF Dye, 1 Vial | 2-8C |
| Part# 4011 | APF Dye, 1 Vial | 2-8C |
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试剂里面含有的是病毒的抗原,现在基本都是人工合成或提取的,跟病毒完全是两回事
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对虾杆状病毒(BP)核酸检测试剂盒(恒温荧光法)
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急性肝胰腺坏死病(AHPND/EMS)病原核酸检测试剂盒(恒温荧光法)
急性肝胰腺坏死病(AHPND/EMS)病原核酸检测试剂盒(PCR-荧光探针法)
对虾偷死野田村病毒(CMNV)核酸检测试剂盒(恒温扩增法)
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它一般在医院、制药企业使用。
试剂盒使用示例: 试剂盒的产生正是为了使实验人员能够摆脱繁重的试剂配制及优化过程,所以试剂盒中一般配备有相应的使用说明书
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