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Mouse IL-4 ELISA Kit (Interleukin 4)

Full Name: Mouse IL-4 ELISA Kit (Interleukin 4)
Reactivity: Mouse
Sample Type: Tissue Homogenates, Plasma, Biological Fluids, Serum
Sensitivity: 9.375 pg/ml

INTRODUCTION

Interleukin 4 functions in the activation of basophils, mast cells and Th2 lymphocytes. It is regarded as an antagonist to the action of IFN-gamma. In T-cells, it is important for the growth and differentiation of Th2 subset, where it promotes the humoral response that is essential external pathogens. In B-cells, it is vital for the stimulation of growth and differentiation. It is also involved in inducing MHC class II and CD23 up-regulation. The primary structure of IL-4 shares 60% homology with human IgE and rat immunoglobulin-like receptor M1 (RIG-I). The initiation and termination codons are identical to those found in IgG1 and IgE respectively, but unlike these proteins, IL4 lacks a hinge region.

The protein encoded by this gene is found in two isoforms due to alternative splicing of the last exon. The shorter, constitutive IL4 isoform lacks the first 126 amino acids of the longer IL4B variant, which encodes a secreted form with a molecular mass of 55 kDa. Interleukin 4 is secreted by proteolysis of an acidic cluster of amino acids at the C-terminus. The resulting 56 kDa molecule is composed of a single polypeptide chain with no disulfide bridges. The shorter isoform thus lacks the first 126 amino acids found in the longer variant, which encodes a secreted form with a molecular mass of 55 kDa. The longer constitutive IL4B variant has been shown to have greater surface expression than the shorter variant, the same observed for the constitutive IL4A variant. The shorter form of IgA may contribute to children with hyper-IgA syndrome, a condition characterized by reduced levels of serum IgA.

Macrophages respond to IL-4 by lowering their production of proinflammatory molecules (TNF and IL-1 beta). This action is responsible for impairing these cells from producing nitrogen or oxygen intermediates, thereby blocking IFN-gamma induced expression of adhesion molecules like E-selectin and ICAM. In addition, macrophages express high levels of both CD40 and TNF receptor II on their surface. These receptors are involved with initiating proinflammatory cytokines when activated by cell surface molecules like LFA-1 integrins or by chemokines. In a study with K562 cells, IL-4 blocked apoptosis and increased cell proliferation.

INTENDED USE

Mouse IL-4 ELISA kit is ideal for analysing concentration of interleukin 4 (IL-4, BSF-1, IL4, BCGF-1) present in mouse samples of tissue homogenates, various biological fluids, plasma and serum.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • One 96-Well Plate: Pre-coated with anti-IL-4 antibody.
  • Standards: Lyophilized recombinant.
  • Sample/Standard Dilution Buffer.
  • Biotinylated-labelled Antibody.
  • Antibody Dilution Buffer.
  • HRP-Streptavidin Conjugate (SABC).
  • SABC Dilution Buffer.
  • TMB Substrate.
  • Wash Buffer (25x).
  • Plate Sealer.
  • Product Instructions.

TYPICAL RESULTS

For this mouse IL-4 ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 15.625, 31.25, 62.5, 125.0, 250.0, 500.0, 1000.0 pg/ml.
Reactivity: Mouse
Sensitivity: 9.375 pg/ml
Range: 14.625 – 1000 pg/ml
Principle: Sandwich
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for IL-4, no cross reactivity or interference between IL-4 and analogues was detected.
– Recovery: Serum (92 – 104%), EDTA Plasma (92 – 100%), Heparin Plasma (90 – 105%).
– Linearity: Serum (86 – 103%), EDTA Plasma (98 – 104%), Heparin Plasma (84 – 100%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

  1. IL-4 enhances survival of in vitro-differentiated mouse basophils through transcription-independent signaling downstream of PI3K. Cell Death Dis. (2018 9 (7): 713. Reinhart R. and Kaufmann T.
  2. Interleukin (IL)-4-independent immunoglobulin class switch to immunoglobulin (Ig)E in the mouse. J Exp Med. (1996) 184 (5): 1651-61. Morawetz R.A., et al.
  3. IL-4-induced STAT6 suppresses IFN-gamma-stimulated STAT1-dependent transcription in mouse macrophages. J Immunol. (1997) 159 (11): 5474-82. Ohmori Y. and Hamilton T.A.
  4. Myeloid cells activate iNKT cells to produce IL-4 in the thymic medulla. Proc Natl Acad Sci U S A. (2019) 116 (44): 22262-22268. Wang H., et al.

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