Standard Curve
Example of Human EGF standard curve in Assay Diluent HB4. Plotted are the background-subtracted data.
This standard curve is provided for demonstration only.
Human EGF ELISA Kit
Human EGF ELISA Kit
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Product Details
Product Details
Product Specification
| Antigen | EGF |
| 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: 4.8%； Inter-assay: 5.1% |
| Sample type | Serum; EDTA Plasma, Heparin Plasma, Citrate Plasma; Cell Culture Supernatant |
| Assay type | Sandwich (quantitative) |
| Sensitivity | 2.18 pg/mL |
| Range | 3.9 pg/mL – 250 pg/mL |
| Recovery | Serum: 106% EDTA Plasma: 108% Citrate plasma: 97% Heparin Plasma: 98% Cell Culture Supernatant: 110% |
| Assay time | 60 minutes |
| Species reactivity | Hu |
Background
Epidermal Growth Factor (EGF) is a potent growth factor that stimulates the proliferation of various epidermal and epithelial cells. Epidermal growth factor (EGF) binds to epidermal growth factor receptor and stimulates an intracellular signal transduction cascade, leading to activation of genes that regulate cell proliferation, angiogenesis, motility, and metastasis. Epidermal growth factor (EGF) mRNA and protein are expressed in a number of adult tissues, especially in epithelial cells in the gastrointestinal tract. Predominant sites of synthesis of this peptide are the submandibular glands, the Brunner glands in the small intestine and the kidney.
Picture
Picture
ELISA
Spike Recovery
The recovery of Human EGF was evaluated in activated samples spiked with concentrations spanning the entire assay range.
Dilution linearity
The concentrations of Human EGF were measured and interpolated from the target standard curves and corrected for sample dilution.
The leftmost bar in the graph represents neat biological samples. "Undiluted" was defined as the neat biological sample (50%). The subsequent bars represent two-fold serial dilutions of this reference. The mean target concentration was determined to be 435.5 pg/mL in human serum.
Spike-and-dilution Linearity
To evaluate assay linearity, samples were spiked with high levels of Human EGF 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 EGF, Mouse EGF and Rat EGF 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 Mouse EGF and Rat EGF were tested in the Human EGF 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.
Serum levels in healthy individuals
Duplicate measurements were performed on serum samples obtained from eight individual healthy donors. Data are reported as dilution factor-corrected interpolated. The resultant mean EGF concentration was 209.2 pg/mL, with a distribution spanning 125.6 to 308.4 pg/mL across the donor cohort.
Protocol Diagram
