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Fluoro ATP

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Fluoro ATP
Fluorescent ADP Detection Kit - Detection of ADP in tissue extracts/cell samples

$345.00$1,595.00

in stock
Product code: FLATP
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Product Description

Introduction

Adenosine Triphosphate (ATP) is an organic molecule which is exclusively formed in the mitochondria. Vritually all living systems store energy in the phosphate bonds of ATP and plays a central  role in cellular metabolism and energy transfer reactions. ATP and be de-phosphorylation to ADP and further de-phosphorylation to AMP. This cycling of ATP is central to transferring potential (thermodynamic) energy from one source to another. There are several genetic disorders that effect the generation of ATP in the mitochondria (1-3).

Cell Technology’s Fluoro ATP assay provides a reliable, sensitive fluorimetric assay for the quantification of ATP in  biological samples.

Key Benefits

  • Detection of ATP in cells or tissue extracts.
  • Detection of ATP in cell death, energy metabolism, mitochondria function.
  • ATP measurement in ATP consuming enzymes such as Kinases and ATPases.
  • ATP detection in Bacterial, Fungal and Plant Cells.
  • Fluorescent 96 well Plate Reader Readout (excitation: 530-570nm and emission at 590-600nm).

Assay Principle

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.

References

  • 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).

Kit contents and Long Term storage

ReagentPart#Temperature
Enzyme Mix 25X6032-20°C
Substrate Buffer3065-20°C
Detection Reagent 100X4028-20°C
ADP Standard 2.5 mM7027-20°C
NEM7026-20°C
x