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> ClinMax™ Multiplex Bead Assay Platform for Flow Cytometry
ClinMax™ Multiplex Bead Assay platform for flow cytometry is an independently developed through rigorously validation to provide multiplexed cytokine detection. The kit is designed to provide a comprehensive solution for preclinical, clinical, and manufacturing quality control by quantifying cytokines using standard curves generated from the same assay, providing a simple tool for drug discovery that is also cost effective.
There are many quantitative detection techniques for soluble proteins (such as cytokines, chemokines, and growth factors), with the most classic being ELISA. However, most of these techniques can only detect a single target per sample. When detecting multiple targets simultaneously, a large sample volume is required, making the process cumbersome and increasing variability between experiments. This is especially challenging in clinical trials, where sample availability is limited and is difficult to perform multiplex or repeat tests.
ClinMax™ Multiplex Bead Assay platform combines flow cytometry with antibody-coated magnetic beads for efficient analyte detection. The kit was developed to detect multiple cytokines utilizing a bead-based sandwich immunoassay. Compared to traditional single-analyte methods like ELISA and Western Blot (WB), it offers significant advantages, including lower sample volume requirements and simpler operation.
| Criteria | Single-plex Method | Multiplex method |
|---|---|---|
Sample volume required | ≥600 uL | 10-50 uL |
Time required | 3-6 days | 0.5 days |
Operation | Simple and straightforward | Complex |
Cost | Low | Cost-effective |
Precision | Low | High |
*For the detection of six soluble proteins
1. Flow cytometry separates magnetic beads based on the fluorescent wavelength emitted by each bead. Specific antibodies are coupled to the surface of each bead which binds to a specific, different molecule.
2. PE-labeled detection antibodies are added to detect the molecules captured on the magnetic bead.
3. PE signal is detected by flow cytometry and quantified against a standard curve.
Principle diagram
Product Advantages
Core Reagents
FAQ
Product Advantages
Low sample volume: < 10 uL per sample.
Fast results: quantitatively evaluate multiple cytokines in a single experiment.
Less steps: easy-to-perform protocol with less steps avoids magnetic bead loss.
Wide detection range: 4 order magnitude standard curve.
Universally compatible: adaptable to various flow cytometers and software.
Protein signal-to-noise ratio was used to screen antibody pairs to ensure performance. Using antibodies with the required sensitivity, cell supernatant, serum, plasma and other samples were used to determine the specific recognition of antibodies to real samples
Magnetic beads are used as the carrier to bind antibodies to the magnetic beads through a chemical reaction. After coupling, antibody coupling amounts were evaluated for consistency between batches and validated.
Non-specific signals from biotin and avidin are reduced by directly labeling by PE.
PBS is used to adjust the concentration to meet the interference level of common interfering substances in the kit samples, such as EDTA and hemoglobin.
ClinMax™ Human Th1/Th2 Cytokine Kit
(Flow Cytometry Multiplex Bead Assay)(Cat. No. FCM-C05R)
ClinMax™ Th1/Th2 Cytokine Kit (Cat. No. FCM-C05R) can quantitatively detect IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ in cell culture medium and serum simultaneously, which greatly improves the work efficiency of researchers and realizes multiplexed analysis of samples.
A representative scatter plot showing single bead population in P1 gate (left), the beads conjugated with analyte-sepcific antibody differntiated by APC fluorescence intensity (middle), and fluorescence signal in propotion to concentration of the analyte was measured for each cytokine in PE channel (right). Data was generated on Beckman Cytoflex S Flow Cytometer, equipped with blue laser (488 nm) and red laser (640 nm).
A double logarithmic fitting curve was plotted for each cytokine (IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ). The present curve of each cytokines was ranging from 9.8 pg/ml to 10000 pg/ml respectively.
The CV of 5 cell culture results with different 3 batches are lower than 20%.（The IL-6 concentrations of cell culture sample II、III are lower than LOQ）
The recovery with 3 different concentrations（2500 pg/ml 、 625 pg/ml 、156.25 pg/ml） spiked into serum, plasma and cell culture are about 80%~120%.
FAQ
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