Fluoro ATP

$345.00$1,595.00

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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.

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#ReagentTemperature
6032Enzyme Mix 25X-20°C
3065Substrate Buffer-20°C
4028Detection Reagent 100X-20°C
7027ADP Standard 2.5 mM-20°C
7026NEM-20°C

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