Fluoro ATP

$345.00$1,595.00

Clear

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).

Additional information

Kit Size

100, 500

Introduction

Adenosine Triphosphate (ATP) is an organic molecule that is exclusively formed in the mitochondria. Virtually 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 affect 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.

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.

Readable Documents

Title Name
msds msds.fluoroATP.pdf
Protocol Fluoro-ATP-Protocol.pdf
Datasheet Fluoro-ATP-Datasheet.pdf

Kit contents and storage

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

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).

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).