Adenosine Assay Kit, IC-6575
Product Introduction
This product is a quantitative assay kit designed for the sensitive and specific measurement of adenosine concentrations in biological samples. Adenosine is a purine nucleoside that functions as a key signaling molecule in numerous physiological and pathological processes, including regulation of immune responses, neurotransmission, cardiovascular function, and cellular energy metabolism. Extracellular adenosine exerts its effects through four G protein-coupled adenosine receptors (A1, A2A, A2B, and A3) and plays critical roles in immunosuppression within the tumor microenvironment, neuroprotection, and modulation of inflammatory responses. This kit provides all necessary reagents to perform a complete adenosine quantification workflow, enabling researchers to accurately measure adenosine levels in cell culture supernatants, tissue homogenates, serum, plasma, and other biological fluids. The assay is suitable for applications in cancer biology, neuroscience, immunology, and cardiovascular research.
Product Features
1. High Sensitivity and Specificity: Detects adenosine with minimal cross-reactivity to structurally related nucleotides and nucleosides including ATP, ADP, AMP, and inosine.
2. Quantitative Measurement: Provides accurate and reproducible adenosine quantification across a linear detection range suitable for typical biological sample concentrations.
3. Complete Kit Format: Includes all necessary reagents, standards, and detailed protocol for a streamlined workflow from sample preparation to data analysis.
4. Broad Sample Compatibility: Validated for use with cell culture supernatants, tissue homogenates, serum, plasma, and other biological fluids.
5. High-Throughput Compatible: 96-well plate format suitable for manual or automated processing in multi-sample studies.
Specifications
Size: 100 assays
Detection Method: Fluorometric or Colorimetric (confirm with specific kit insert)
Format: 96-well plate
Sample Types: Cell culture supernatants, tissue homogenates, serum, plasma, biological fluids
Storage and Stability
Storage Conditions: Store kit components at -20¡ãC or 4¡ãC as indicated on individual component labels. Protect 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: Read the entire protocol before beginning. Protect light-sensitive reagents from exposure. Prepare all working solutions freshly before use.
1. Sample Preparation: Collect biological samples and process as appropriate. For cell culture supernatants, centrifuge at 1,000 x g for 5 minutes to remove debris. For tissue samples, homogenize in PBS or appropriate buffer, centrifuge at 10,000 x g for 10 minutes at 4¡ãC, and collect the supernatant. For serum or plasma, use samples directly or deproteinize as recommended in the detailed protocol.
2. Standard Curve Preparation: Prepare adenosine standard solutions by serial dilution of the provided adenosine standard stock in assay buffer to generate a standard curve typically covering the range of 0.1-100 µM or as specified in the lot-specific datasheet.
3. Assay Setup: Add standards, samples, and appropriate controls (50-100 µL per well) to the designated wells of the provided 96-well assay plate according to the plate layout plan. Add reaction mix containing adenosine-detecting enzyme(s), probe, and cofactors as directed.
4. Incubation: Incubate the plate at room temperature or 37¡ãC for 30-60 minutes protected from light, according to the specific protocol for the detection method selected.
5. Detection and Analysis: Measure the absorbance or fluorescence signal using a microplate reader at the appropriate wavelengths specified in the protocol. Calculate adenosine concentrations from the standard curve, correcting for any sample dilution factors applied during preparation.
Precautions
1. Protect reagents from light during handling and storage; light exposure may degrade light-sensitive components and reduce assay sensitivity.
2. Avoid repeated freeze-thaw cycles of reconstituted reagents; aliquot reconstituted enzyme and probe solutions into single-use volumes and store at -80¡ãC.
3. Use appropriate sample controls including blank wells and known adenosine-spiked samples to validate assay performance with each experiment.
4. Certain biological samples may contain substances that interfere with the assay; deproteinization or sample cleanup steps may be required for complex matrices.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What is the detection principle of this assay?
A1: The assay employs an enzymatic cascade reaction that specifically converts adenosine to a quantifiable product, generating a fluorescent or colorimetric signal proportional to the adenosine concentration in the sample. Specific enzyme and probe details are provided in the kit insert.
Q2: What is the linear detection range of the assay?
A2: The assay typically provides a linear detection range of 0.1-100 µM adenosine. Consult the lot-specific datasheet for the exact standard curve parameters and detection limit information.
Q3: How should I prepare tissue samples for adenosine measurement?
A3: Tissues should be rapidly homogenized in cold PBS or assay buffer containing adenosine deaminase inhibitors, centrifuged at 10,000 x g for 10 minutes at 4¡ãC to remove debris, and the supernatant used immediately or stored at -80¡ãC. Avoid multiple freeze-thaw cycles as adenosine is rapidly metabolized.
Q4: What is the specificity of the assay for adenosine versus other nucleotides?
A4: The assay exhibits high specificity for adenosine with minimal cross-reactivity (<1%) with ATP, ADP, AMP, inosine, and other structurally related nucleosides. However, verify specificity for your specific sample matrix using appropriate controls.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of sample preparation and assay conditions is recommended for specific sample types and experimental systems.
3. 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.
4. Wear appropriate protective clothing and gloves when handling this product and biological samples.
Ordering Information
Catalog Number: IC-6575
Product Name: Adenosine Assay Kit
Size: 100 assays
Price: CNY ¥7550.00 / USD $755.00 / EUR €906.00 / JPY ¥135900.00
2023 Version
