Rat IL-4 serum & plasma OneStep ELISA Kit

Catalog Number: S0C3245 Reactivity: Rat Conjugation: Brand: Starter

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$170 USD

Size: 1x24T 1x48T 1x96T 5x96T 10x96T

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Product Details

Product Specification

Antigen	IL-4
Immunogen	Recombinant Protein
Antibody Type	Recombinant mAb
Reactivity	Rat
Purification	Protein A
Stability & Storage	12 months from date of receipt / reconstitution, 2 to 8°C as supplied.

Kit

Precision	Intra-assay: 3.4% Inter-assay: 4.1%
Sample type	Serum; Plasma
Assay type	Sandwich (quantitative)
Sensitivity	3.24 pg/mL
Range	18.8 pg/mL – 1200 pg/mL
Recovery	SD Rat Serum: 109% SD Rat EDTA Plasma: 108% SD Rat heparin Plasma: 101% Lewis Rat Serum: 96% Lewis Rat EDTA Plasma: 96% SHR Rat EDTA Plasma: 101% SHR Rat citrate Plasma: 99%
Assay time	60 minutes

Background

Rat interleukin-4 (IL-4) is a pleiotropic cytokine primarily produced by activated T cells, mast cells, basophils, and NK1.1+ T cells. It plays a central role in driving Th2 immune responses and is a key regulator of B-cell activation, inducing class II MHC expression and enhancing the secretion of IgE and IgG1. The protein signals through the interleukin-4 receptor, activating the JAK/STAT6 pathway, which mediates its immunoregulatory functions. The full-length rat IL-4 precursor is 147 amino acids with a predicted mass of approximately 16.2 kDa. The mature secreted protein, typically produced recombinantly as a 126-amino-acid globular polypeptide, has a molecular weight of about 14.0 kDa. Recombinant rat IL-4 is commonly expressed in E. coli and its biological activity is often confirmed by its ability to stimulate the proliferation of rat splenocytes. Given its critical role in Th2-mediated immunity and allergic responses, rat IL-4 is an essential tool for studying these pathways in preclinical models.

Picture

ELISA

Standard curve
Example of Rat IL-4 standard curve in Assay Diluent TA11.

Spike Recovery
The recovery of Rat IL-4 was evaluated in activated samples spiked with concentrations spanning the entire assay range.

Cross-reactivity
The heatmap displays OD450 ratios for serum samples from multiple species. Color intensity is proportional to the O.D.450 ratio (test species / reference species). Darker colors indicate higher cross-reactivity.

Spike-and-dilution Linearity
To evaluate assay linearity, three samples were spiked with high levels of Rat IL-4 in different matrices and serially diluted with the corresponding Calibrator Diluent to fall within the assay's dynamic range.

Cross-reactivity
We evaluated cross-reactivity by testing human IL-4, mouse IL-4, monkey IL-4, and rat IL-13 in the same assay system. Each protein was measured in a series of dilutions, and the dose-response curves were compared. When rat IL-4 was tested in assays for human IL-4, mouse IL-4, monkey IL-4, and rat IL-13, no significant cross-reactivity was observed, as the signal stayed within the background range of the assays.

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

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.

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.