Monkey Th1/Th2 6-Plex Kit (Flow Cytometry Multiplex Bead Assay)

CBA

• 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.

• Distribution Diagram
  The figure shows the distribution of microsphere clusters corresponding to different cytokines; it enables the simultaneous differentiation of multiple cytokines, providing a basis for the group definition for the subsequent quantitative analysis of each cytokine.

• Distribution Diagram
  The histogram reflects the number of particles in each microsphere group.

• Standard curve
  Example of IL-2, IL-4, IL-6, IL-10, IFN-γ, TNF-α standard curve in Assay Diluent .

• Quantifiction of Monkey PBMC
  PBMC Unsimulated Supernatant: The cells of PBMCs were cultured for 24 hours, the cell culture supernatant was collected and analyzed quantitatively using S0Q1004.
  PBMC simulated Supernatant: The cells of PBMCs were stimulated with 10 ng/mL PMA. After 24 hours, the cell culture supernatant was collected and analyzed quantitatively using S0Q1004.

• Linearity
  After serial dilution of the high-concentration combined proteins of monkey IL-2, IL-4, IL-6, IL-10, IFN-γ, TNF-α using plasma, serum, and cell culture supernatant matrices respectively, the concentrations of Monkey IL-2, IL-4, IL-6, IL-10, IFN-γ, TNF-α 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 IL-2, IL-4, IL-6, IL-10, IFN-γ, TNF-α were tested.

• Inter-assay precision
  Ten repeated tests were conducted on three different levels of IL-2, IL-4, IL-6, IL-10, IFN-γ, TNF-α using different batches of reagents.

• Specificity
  The antibodies used in the kit have been screened for their specific reactivity with particular cytokines. After analyzing samples containing only a single recombinant cytokine protein, it was found that when using this detection method, there was no cross-reaction or background detection of cytokines 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 cytokine 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 cytokine proteins.
  The results were compared with those of samples that had the same concentration of cytokines added in the assay buffer.

• Protocol Diagram