Standard curve
Example of Monkey IL-4 standard curve in Assay Diluent HR2.
Monkey IL-4 Serum & Plasma OneStep ELISA Kit
Monkey IL-4 Serum & Plasma OneStep ELISA Kit
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Product Details
Product Details
Product Specification
| Antigen | IL-4 |
| Immunogen | Recombinant Protein |
| Antibody Type | Recombinant mAb |
| Reactivity | Mk |
| Purification | Protein A |
| Stability & Storage | 12 months from date of receipt / reconstitution, 2 to 8°C as supplied. |
Kit
| Precision | Intra-assay: 1.8% Inter-assay: 2.1% |
| Sample type | Serum; Plasma |
| Assay type | Sandwich (quantitative) |
| Sensitivity | 0.190 pg/mL |
| Range | 3.1 pg/mL – 200 pg/mL |
| Recovery | Monkey Serum: 107% EDTA Plasma: 113% |
| Assay time | 60 minutes |
| Species reactivity | Hu |
Background
Monkey Interleukin-4 (IL-4) is a pleiotropic cytokine primarily produced by activated T cells, mast cells, and bone marrow stromal cells, playing a key role in regulating immune responses. It has diverse biological functions, including stimulating the proliferation of activated B-cells and T-cells, and promoting the differentiation of naive CD4+ T-cells into Th2 cells. IL-4 also enhances the secretion and cell surface expression of IgE and IgG1, and regulates the low-affinity Fc receptor for IgE (CD23) on lymphocytes and monocytes, which is central to allergic and anti-parasitic responses. The protein exerts its effects by binding to the interleukin-4 receptor, a complex that can also bind IL-13, explaining their overlapping functions; this binding activates the STAT6 signaling pathway to mediate its immunoregulatory signals. Recombinant monkey IL-4 proteins, derived from cynomolgus or rhesus macaques, are typically produced in systems like E. coli, yeast, or HEK293 cells for research applications. These recombinant proteins generally consist of approximately 129 amino acids with a molecular weight around 14.9 kDa, though glycosylation in mammalian systems can result in higher apparent molecular weights. Due to the high immunological similarity between monkeys and humans, monkey IL-4 serves as an essential tool for studying Th2-mediated immune pathways, allergic diseases, and related therapeutic strategies in non-human primate models.
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ELISA
Spike Recovery
The recovery of Monkey IL-4 was evaluated in activated samples spiked with concentrations spanning the entire assay range.
Spike-and-dilution Linearity
To evaluate assay linearity, three samples were spiked with high levels of Monkey IL-4 in different matrices and serially diluted with the corresponding Calibrator Diluent to fall within the assay's dynamic range.
Intra-Assay Precision (Precision within an assay)
Three samples of known concentration were tested fifteen times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays).
Three samples of known concentration were tested in separate assays to assess inter-assay precision. Assays were performed with at least three lots of components.
Determination of Minimum Detectable Dose (MDD)
The MDD was determined using three independent lots of assay components. For each lot, 19 replicate measurements of the diluent (zero calibrator) were performed. The mean (AVERAGE) and standard deviation (STDEV) of the 19 replicates were calculated. The MDD for each lot was then calculated according to the following formula:
MDD = 2 × STDEV + AVERAGE
Cross-reactivity
The cross-reactivity among Mouse IL-4, Monkey IL-4, and Human IL-4 was evaluated by testing them in the same assay system. Serial dilutions of each protein were measured, and the dose-response curves were compared. When Mouse IL-4 was tested in the Monkey IL-4 assay, no significant cross-reactivity was observed, and the signal remained within the background range of the assay. When Human IL-4 was tested in the Monkey IL-4 assay, significant cross-reactivity was observed.
HOOK Effect Threshold
The upper limit of the HOOK effect was established at 100× the highest calibrator concentration (equivalent to a 2-log10 increase). No HOOK effect was observed below this threshold, confirming that samples within this concentration range yield reliable quantitative results without signal depression.
Protocol Diagram
