Human IL-4 ELISA Kit

Protocol Booklet

Key features and details

Specification: 96 Test
Sensitivity: 0.03 pg/ml (50 μl) ；0.28 pg/ml (10 μl)
Standard Curve Range: 1.37~1000 pg/ml
Standard Curve Gradient: 7 Points/3 Folds
Number of Incubations: 2
Sample Volume: 50 μl/10 μl
Assay type: Sandwich Elisa
Operation Duration: 120min

Brand:

CAT.NO. : AEH0007

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Size:

96T

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Product Details

Background

Interleukin-4 (IL-4), also known as B cell-stimulatory factor-1, is a monomeric, approximately 13 kDa ‑ 18 kDaTh2 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.

Typical data

pg/ml	O.D.		Average	Corrected
0	0.0089	0.0085	0.0087
1.37	0.0163	0.0168	0.0166	0.0079
4.12	0.031	0.0371	0.0341	0.0254
12.35	0.0855	0.0907	0.0881	0.0794
37.04	0.2498	0.2553	0.2526	0.2439
111.11	0.7143	0.7335	0.7239	0.7152
333.33	1.9249	1.9264	1.9257	1.917
1000	3.8591	3.7755	3.8173	3.8086

Precision

	Intra-assay Precision			Inter-assay Precision
Sample Number	S1	S2	S3	S1	S2	S3
Sample Number	22	22	22	6	6	6
Average(pg/ml)	24.9	124.6	243.4	28.5	126.1	219.8
Standard Deviation	1.3	5.5	8.2	1.9	6	6.4
Coefficient of Variation(%)	3.4	4.5	5.1	6.5	4.7	2.9

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 human IL-4 into health human serum sample. The un-spiked serum was used as blank in this experiment.

The recovery ranged from 88% to 105% with an overall mean recovery of 98%.

Sample Values

Sample Matrix	Sample Evaluated	Range (pg/ml)	Detectable (%)	Mean of Detectable (pg/ml)
Serum	30	n.d.-0.49	80	0.13

Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of IL-10 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.

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