Standard Curve
Example of Human IL-3 standard curve in Assay Diluent TA2. Plotted are the background-subtracted data.
This standard curve is provided for demonstration only.
Human IL-3 OneStep ELISA Kit
Human IL-3 OneStep ELISA Kit
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Product Details
Product Details
Product Specification
| Antigen | IL-3 |
| Immunogen | Recombinant Protein |
| Antibody Type | Recombinant mAb |
| Reactivity | Hu |
| Purification | Protein A |
| Stability & Storage | 12 months from date of receipt / reconstitution, 2 to 8°C as supplied. |
Kit
| Precision | Intra-assay: 7.6%； Inter-assay: 9.8% |
| Sample type | Serum; EDTA Plasma, Heparin Plasma, Citrate Plasma; Cell Culture Supernatant |
| Assay type | Sandwich (quantitative) |
| Sensitivity | 115.81 pg/mL |
| Range | 312.5 pg/mL – 20,000 pg/mL |
| Recovery | Serum: 111% EDTA Plasma: 117% Citrate plasma: 93% Heparin Plasma: 104% Cell Culture Supernatant: 111% |
| Assay time | 60 minutes |
| Species reactivity | Hu |
Background
Interleukin-3 (IL-3) is a secreted cytokine that stimulates the proliferation and differentiation of pluripotent hematopoietic stem cells and various lineage-committed progenitors. It also regulates the activation of mature mast cells, basophils, eosinophils, and macrophages. It is produced by activated T cells, thymic epithelial cells, activated mast cells, keratinocytes, neurons, and astrocytes. IL-3 signals through a receptor complex that contains IL-3 R alpha/CD123 and the common beta chain which is also a component of the receptors for IL-5 and GM-CSF. Receptors for IL-3 are present on bone marrow progenitors, macrophages, mast cells, eosinophils, megakaryocytes, basophils, and various myeloid leukemic cells. Human IL-3 does not show activity on mouse cells.
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ELISA
Spike Recovery
The recovery of Human IL-3 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 Human IL-3 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 between Human IL-3 and Rat LI-3 was assessed by testing both proteins in the same assay system. Serial dilutions of each protein were measured, and the dose-response curves were compared. No significant cross-reactivity was observed when Human NT-pro BNP was tested in the Human IL-3 BNP assay, as the signals remained within the background range of the assay.
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
