ADP/ATP Ratio BioAssay™ Kit

Cat# 298992-100T

Size : 100Tests

Brand : US Biological



Shipping Temp
Dry Ice

Storage Temp
-20°C

Changes in the ADP/ATP ratio have been used to differentiate modes of cell death and viability. Increased levels of ATP and decreased levels of ADP signify proliferating cells. Conversely, decreased levels of ATP and increased levels of ADP represent apoptotic or necrotic cells where the decrease in ATP and increase in ADP is much more pronounced in necrosis versus apoptosis.

Sample Type:
Mammalian cells

Intended Use:
Apoptosis and Necrosis determination: in cells.
Cell proliferation: effects of cytokines, growth factor, nutrients.
Drug discovery: high-throughput screening for anticancer drugs.

Test Principle:
This ADP/ATP Ratio BioAssay™ Kit provides a rapid method to measure ADP and ATP levels for the screening of apoptosis, necrosis and cell proliferation in mammalian cells. The assay involves two steps. In the first step, the working reagent lyses cells to release ATP and ADP. In the presence of luciferase, ATP immediately reacts with the Substrate D-luciferin to produce light. The light intensity is a direct measure of intracellular ATP concentration.
Luciferase
ATP + D-luciferin + O2 oxyluciferin + AMP + PPi + CO2 + light
In the second step, the ADP is converted to ATP through an enzyme reaction. This newly formed ATP then reacts with the D-luciferin as in the first step. This non-radioactive, homogeneous cell-based assay is performed in microplates. The reagent is compatible with all culture media and with all liquid handling systems for high-throughput screening applications in 96- well and 384-well plates.

Kit Components:
Assay Buffer, 1x10ml
Substrate, 1x120ul
Cosubstrate, 1x120ul
ATP Enzyme, 1x120ul
ADP Enzyme, 1x120ul

Storage and Stability:
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.