ATP Assay Kit, IC-6560
Product Introduction
This product is a bioluminescence-based assay kit for the quantitative measurement of adenosine triphosphate (ATP) in biological samples. ATP is the universal energy currency of all living cells, serving as the primary high-energy phosphate donor for numerous energy-dependent cellular processes including biosynthesis, active transport, signal transduction, and cell motility. Cellular ATP levels are tightly regulated and reflect the balance between ATP synthesis (primarily through oxidative phosphorylation and glycolysis) and ATP consumption. Depletion of ATP is a hallmark of apoptosis, necrosis, and metabolic dysfunction, making ATP quantification a valuable indicator of cell viability, proliferation, and metabolic health. This kit employs the highly specific firefly luciferase-catalyzed reaction, in which ATP drives the oxidation of D-luciferin to oxyluciferin, generating bioluminescence at 560 nm. The luminescence signal is directly proportional to ATP concentration over a wide linear range, providing exceptional sensitivity and specificity. The kit is compatible with cell lysates, tissue homogenates, and biological fluids.
Product Features
1. Ultra-Sensitive Bioluminescent Detection: Firefly luciferase-based assay provides femtomolar sensitivity for ATP detection with excellent signal-to-noise ratios.
2. Exceptional Specificity: Luciferase reaction is strictly ATP-dependent, with negligible cross-reactivity with other nucleotide triphosphates.
3. Broad Linear Range: Linear detection over several orders of magnitude, accommodating diverse sample types and ATP concentrations.
4. Simple and Rapid Protocol: Single-step "add-and-read" procedure with luminescence detection within minutes.
5. Complete Kit Format: Includes all necessary enzymes, substrates, ATP standard, and lysis buffer for a complete ATP assay workflow.
Specifications
Size: Available in 100 assays and 200 assays
Detection Method: Bioluminescence (560 nm)
Format: 96-well plate (white or black opaque plates recommended)
Sample Types: Cell lysates, tissue homogenates, biological fluids
Storage and Stability
Storage Conditions: Store kit components at -20¡ãC, protected from light. The luciferase/luciferin reagent is particularly sensitive and must be stored protected from light.
Shelf Life: The product is stable for 12 months from the date of manufacture when stored as directed. Avoid repeated freeze-thaw cycles of reconstituted reagents.
Protocol (For Reference Only)
Important: Use white or black opaque 96-well plates for luminescence detection. Protect luciferase/luciferin reagent from light. Prepare all working solutions freshly before use.
1. Sample Preparation: For cultured cells, aspirate culture medium, wash once with PBS, and add ATP Lysis Buffer directly to cells. Incubate at room temperature for 5 minutes with gentle shaking. Collect lysate and centrifuge at 12,000 x g for 5 minutes at 4¡ãC. Transfer supernatant to a clean tube. For tissue homogenates, homogenize tissue in ATP Lysis Buffer, centrifuge at 12,000 x g for 10 minutes at 4¡ãC, and collect supernatant.
2. Standard Curve Preparation: Prepare a serial dilution of the provided ATP standard in ATP Lysis Buffer to generate a standard curve typically covering 1 nM to 10 µM. Include a blank well containing lysis buffer only.
3. Reaction Setup: Transfer 50-100 µL of each standard, diluted sample, and blank to designated wells of a white 96-well plate. Prepare Luciferase/Luciferin Working Solution as directed in the protocol. Add an equal volume of Working Solution to each well.
4. Detection: Mix the plate briefly and measure luminescence immediately using a microplate reader capable of luminescence detection. Use an integration time of 1-5 seconds per well.
5. Data Analysis: Subtract blank values from all readings. Generate a standard curve by plotting luminescence (RLU) versus ATP concentration, and calculate ATP concentrations in unknown samples by interpolation.
Precautions
1. Protect luciferase/luciferin reagent from light; light exposure reduces enzymatic activity and assay sensitivity.
2. ATP is rapidly consumed by cellular ATPases; process samples promptly and maintain on ice to preserve ATP levels.
3. Use white or black opaque plates to minimize well-to-well crosstalk during luminescence detection.
4. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What is the detection limit of this assay?
A1: The assay can detect ATP concentrations as low as 0.1-1 nM (fmol range per well), depending on the luminometer sensitivity and sample preparation. This sensitivity is sufficient for detecting ATP from as few as 10-100 cells.
Q2: Can I measure ATP in tissue samples?
A2: Yes. Rapidly homogenize tissue in ice-cold ATP Lysis Buffer using a homogenizer. Centrifuge to remove debris and assay the supernatant immediately. Normalize ATP content to tissue weight or protein concentration.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
3. Wear appropriate protective clothing and gloves when handling this product.
Ordering Information
Catalog Number: IC-6560
Product Name: ATP Assay Kit
Size: 100 assays / 200 assays
Price:
100 assays: CNY ¥1530.00 / USD $153.00 / EUR €183.60 / JPY ¥27540.00
200 assays: CNY ¥2450.00 / USD $245.00 / EUR €294.00 / JPY ¥44100.00
