Rat IL-4 enzyme-linked immunoassay kit

CAT: ER0003 Datasheet
Specification 96 Test
Sensitivity 17.29 pg/ml (50 μl)
Standard Curve Range 2.74~2000 pg/ml
Standard Curve Gradient 7 Points/3 Folds
Number of Incubations 2
Sample Volume 50 μl
Type Fully Ready-to-Use
Operation Duration 120min
pg/ml O.D. Average Corrected
0 0.105 0.104 0.1045
2.74 0.1086 0.1036 0.1061 0.0016
8.23 0.11 0.108 0.109 0.0045
24.69 0.1382 0.1433 0.1408 0.0767
74.07 0.2812 0.2953 0.2883 0.2242
222.22 0.6422 0.6543 0.6483 0.5842
666.67 1.5432 1.5153 1.5293 1.4652
2000 3.0881 3.1932 3.1407 3.0766

Precision

Intra-assay Precision Inter-assay Precision
Sample Number S1 S2 S3 S1 S2 S3
22 22 22 6 6 6
Average(pg/ml) 33.4 172.8 557.4 32.8 175.6 561.2
Standard Deviation 1.8 8.1 26.8 2.3 12.1 34.8
Coefficient of Variation(%) 5.4 4.7 4.8 7 6.9 6.2

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays) Three samples of known concentration were tested six times on one plate to assess intra-assay precision.

Spike Recovery

The spike recovery was evaluated by spiking 3 levels of rat IL-4 into health rat serum sample. The un-spiked serum was used as blank in these experiments.

The recovery ranged from 93% to 115% with an overall mean recovery of 105%.

Sample Values

Sample Matrix Sample Evaluated Range (pg/ml) Detectable (%) Mean of Detectable (pg/ml)
Serum 30 n.d. 0 0

Serum/Plasma – Thirty samples from apparently healthy rats were evaluated for the presence of IL-6 in this assay. No medical histories were available for the donors.

n.d. = non-detectable. Samples measured below the sensitivity are considered to be non-detectable.

Background: IL-4

Interleukin-4 (IL-4), also known as B cell-stimulatory factor-1, is a monomeric, approximately 13 kDa ‑ 18 kDa Th2 cytokine that shows pleiotropic effects during immune responses. It is a glycosylated polypeptide that contains three intrachain disulfide bridges and adopts a bundled four alpha -helix structure. Human IL-4 is synthesized with a 24 aa signal sequence. Alternate splicing generates an isoform with a 16 aa internal deletion. Mature human IL-4 shares 55%, 39% and 43% aa sequence identity with bovine, mouse, and rat IL-4, respectively. Human, mouse, and rat IL-4 are species-specific in their activities.

IL-4 exerts its effects through two receptor complexes. The type I receptor, which is expressed on hematopoietic cells, is a heterodimer of the ligand binding IL-4 R alpha and the common gamma chain (a shared subunit of the receptors for IL-2, -7, -9, -15, and ‑21). The type II receptor on nonhematopoietic cells consists of IL-4 R alpha and IL-13 R alpha 1. The type II receptor also transduces IL-13 mediated signals. IL-4 is primarily expressed by Th2-biased CD4+ T cells, mast cells, basophils, and eosinophils. It promotes cell proliferation, survival, and immunoglobulin class switch to IgG4 and IgE in human B cells, acquisition of the Th2 phenotype by naïve CD4+ T cells, priming and chemotaxis of mast cells, eosinophils, and basophils, and the proliferation and activation of epithelial cells. IL-4 plays a dominant role in the development of allergic inflammation and asthma.

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