Distribution Diagram
In the figure, the R1 gate represents the selected target capture microsphere group, which is used to define this group for subsequent detection and analysis of fluorescence signals.
Human Essential Immune Biomarker 7-Plex Kit (Flow Cytometry Multiplex Bead Assay)
Human Essential Immune Biomarker 7-Plex Kit (Flow Cytometry Multiplex Bead Assay)
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Product Details
Product Details
Product Specification
| Antigen | IL-4、IL-6、IL-8、IL-10、IFN-γ、TNF-α、Granzyme B |
| Reactivity | Human |
| Stability & Storage | 12 months from date of receipt / reconstitution, 2 to 8°C as supplied. |
Kit
| Precision | Intra-assay: ＜10%； Inter-assay: ＜15% |
| Sample type | Serum; Plasma-EDTA; Plasma-Heparin; Plasma-Citrate; Cell Culture Supernatant |
| Assay type | Sandwich (quantitative) |
| Sensitivity | ＜6.0 pg/mL |
| Recovery | 70% - 130% |
| Assay time | 70 minutes |
| Species reactivity | Human |
| CROSS REACTIVITY | <1% |
Background
The Cytometric Bead Array (CBA) technology enables the capture of soluble analytes or analyte panels using beads with known sizes and fluorescence intensities, thereby allowing the detection of analytes via flow cytometry.
Each capture bead in the CBA kit is conjugated with a specific antibody. The detection reagent provided with the kit is Streptavidin-Phycoerythrin, and the intensity of the fluorescent signal generated by this reagent is proportional to the amount of bound analyte.
When the capture beads and detection reagent are incubated together with the test sample containing target analytes, sandwich complexes are formed. Detection of these complexes by flow cytometry allows for the identification of particles exhibiting the fluorescence characteristics of both the beads and the detection reagent.
Picture
Picture
CBA
Distribution Diagram
The figure shows the distribution of microsphere clusters corresponding to different biomakers; it enables the simultaneous differentiation of multiple biomakers, providing a basis for the group definition for the subsequent quantitative analysis of each biomaker.
Distribution Diagram
The histogram reflects the number of particles in each microsphere group.
Distribution Diagram
This figure shows a typical flow cytometry scatter plot of the CBA quantitative detection reagent. The captured microspheres with different APC fluorescence intensities are sequentially enclosed (A1 to A2 and A4 to A8), corresponding to the 7 different target substances detectable in the kit; the signal intensity of the PE channel is proportional to the content of the target substances bound to each microsphere.
Standard curve
Example of IL-4, IL-6, IL-8, IL-10, IFN-γ, TNF-α, Granzyme B standard curve in Assay Diluent .
Quantifiction of Human PBMC
PBMC Unsimulated Supernatant: The cells of PBMCs were cultured for16 hours, the cell culture supernatant was collected and analyzed quantitatively using S0Q1007.
PBMC simulated Supernatant: The cells of PBMCs were stimulated with 100 ng/mL LPS 10 ng/mL IL-1α. After 16 hours, the cell culture supernatant was collected and analyzed quantitatively using S0Q1007.
Linearity
After serial dilution of the high-concentration combined proteins of Human IL-4, IL-6, IL-8, IL-10, IFN-γ, TNF-α, Granzyme B using plasma, serum, and cell culture supernatant matrices respectively, the concentrations of Human IL-4, IL-6, IL-8, IL-10, IFN-γ, TNF-α, Granzyme B were measured and interpolated from the target standard curves and corrected for sample dilution.
Intra-assay precision
Ten replicates of each of three different levels of human IL-4, IL-6, IL-8, IL-10, IFN-γ, TNF-α and Granzyme B were tested.
Inter-assay precision
Ten repeated tests were conducted on three different levels of human IL-4, IL-6, IL-8, IL-10, IFN-γ, TNF-α and Granzyme B using different batches of reagents.
Specificity
The antibodies used in the kit have been screened for their specific reactivity with particular biomarkers. After analyzing samples containing only a single recombinant biomaker protein, it was found that when using this detection method, there was no cross-reaction or background detection of biomarkers in other capture magnetic bead groups.
Theoretical limit of detection
Repeat the detection of the blank sample 20 times using the same batch of reagents, and calculate the theoretical limit of detection.
Recovery
Within the detection range, the combined biomaker proteins were added to different matrices at high, medium, low concentrations.
The matrices used in these experiments were diluted 2-fold with the assay buffer before the addition of the biomaker proteins.
The results were compared with those of samples that had the same concentration of biomakers added in the assay buffer.
Protocol Diagram
