- Description
- Additional Information
- Readable Documents
- Assay Principle
- Reviews
Additional information
| Kit Size | 100, 500 |
|---|
The Fluoro ATP detection kit utilizes a non-fluorescent detection reagent, which is reduced in the presence ATP and a coupled enzyme reaction to produce its fluorescent analog. There is a linear relationship of ATP concentration to the fluorescent analog concentration. An ATP standard curve is generated to interpolate sample ATP concentrations. The kit can be used in both endpoint and kenitic modes.
Reaction:
1. ATP + Enzyme Coupled Reaction + Non-Fluorescent Detection Reagent → Fluorescent Analog + AMP
Detection: Excitation: 530-570nm and Emission at 590-600nm
2. 50µL of sample or ATP Standard
+
50µL of Enzyme Reaction Cocktail
↓
Incubate 30 – 60 minutes; RT; DARK
Read on Plate Reader: Excitation: 530-570nm
Emission at 590-600nm
Figure 1. ATP vs ADP Standard Curve fitted with linear regression.
ATP Spike | % Recovery | % Recovery |
no NEM | 40mM NEM | |
25.5 | 170 | 95.85 |
8.35 | 230 | 102 |
| 2.154 | 227 | 86.75 |
Table 1. ATP was spiked into Jurkat cell samples in Substrate Buffer (Part# 3046) with or without 40mM NEM (Part# 7026). % Recovery was determined via linear regression from ATP standard curve. N=3 per sample.
| Document Title |
| Fluoro ATP Protocol |
| Fluoro ATP Datasheet |
| msds.fluoroATP |
| Reference |
| Mitchell, P., Coupling of phosphorylation to electron and hydrogen transfer by a chemi-osmotic type of mechanism. Nature, 191, 144–148 (1961). |
| Alirol, E., and Martinou, J.C., Mitochondria and cancer: is there a morphological connection? Oncogene, 25, 4706–4716 (2006). |
| Carrozzo, R. et al., Infantile mitochondrial disorders. Biosci. Rep., 27, 105–112 (2007). |
| Part# | Reagent | Temperature |
| 6032 | Enzyme Mix 25X | -20°C |
| 3065 | Substrate Buffer | -20°C |
| 4028 | Detection Reagent 100X | -20°C |
| 7027 | ADP Standard 2.5 mM | -20°C |
| 7026 | NEM | -20°C |
