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IL-4R alpha/CD124 Protein, Human (HEK293)

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IL-4R alpha is a subunit alpha shared by IL-4 and IL-13 receptors, found in leukocytes originally. IL-4R alpha couples to the JAK1/2/3-STAT6 pathway and involves in promoting Th2 differentiation. IL-4R alpha/CD124, Human consists of 825 amino acids (M1-S825) with a transmembrane domain (233-256 a.a) and a soluble form (1-227 a.a). Soluble IL-4R (sIL-4R) inhibits IL4-mediated cell proliferation and IL-5 up-regulation by T-cells. IL-4R alpha/CD124, Human (G24-H232) is produced in HEK293 cells with a full length of 209 amino acids.

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  • Species: Human
  • Source: HEK293
  • Storage:
    Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.
  • Experimental Validation Results for IL-4R alpha/CD124 Protein, Human (HEK293)
  • Experimental Validation Results for IL-4R alpha/CD124 Protein, Human (HEK293)
  • IL-4R alpha/CD124 Protein, Human (HEK293) Product Image
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  • Biological Activity
  • Technical Parameters
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Biological Activity

Description

IL-4R alpha is a subunit alpha shared by IL-4 and IL-13 receptors, found in leukocytes originally. IL-4R alpha couples to the JAK1/2/3-STAT6 pathway and involves in promoting Th2 differentiation[1]. IL-4R alpha/CD124, Human consists of 825 amino acids (M1-S825) with a transmembrane domain (233-256 a.a) and a soluble form (1-227 a.a). Soluble IL-4R (sIL-4R) inhibits IL4-mediated cell proliferation and IL-5 up-regulation by T-cells[1]. IL-4R alpha/CD124, Human (G24-H232) is produced in HEK293 cells with a full length of 209 amino acids.

Background

Interleukin-4R alpha (IL-4Rα), also known as CD124 and B cell stimulatory factor (BSF) receptor, is one of the anti-inflammatory cytokines, and highly expressed in activated T-cells[1].
IL-4R alpha participates in forming two interleukin receptors in different cell types. For the type I receptor, depends on IL-4R alpha binding IL-4 to recruit IL-2R gamma chain in immune cells. IL-2R gamma is the common subunit for a variety of interleukin receptors, involved in the stimulation of neutrophil phagocytosis by IL-15. For the type II receptor, depends on IL-4R alpha binding IL-4 to recruit IL-13R alpha 1 chain. IL-13R alpha 1 is an alternat accessory protein to the common cytokine receptor gamma chain in non-immune cells[2][3].
The sequence of amino acids in IL-4R alpha proteins in human is very different from mouse (53.35%), or rat (52.82%).
IL-4 R alpha generates a soluble form by alternate splicing or proteolysis, maintaining ligand binding properties and inhibiting IL-4 bioactivity. IL-4 R alpha soluble isoform 1 can be produced by proteolytic cleavage at the cell surface (shedding) by a metalloproteinase[4].
IL-4 R alpha plays an important role in Th2-biased immune responses, alternative macrophage activation, mucosal immunity, allergic inflammation, tumor progression, and atherogenesis[5].

In Vitro

Interleukin-4Rα (IL-4Rα) shows promotion of Th2 cytokine and IgE response without hIL-4, and fails to expel N. brasiliensis worms in transgenic mice (hIL-4RαTg/mIL-4Rα-/-) infected with Nippostrongylus brasiliensis[8].

In Vivo

Interleukin-4Rα (IL-4Rα) contains a immunoreceptor tyrosine-based inhibitory motifs (ITIM), plays a functional role in the regulation of IL-4-induced proliferation, ablation of ITIM results in a hyperproliferative response to IL-4 stimulation in 32D/IRS-2 cells expressing mutant IL-4R α-chains (△712 and Y713F)[6].
Recombinant sIL-4R (10 ng/mL; 3 d) inhibits IL-4-mediated proliferation and IL-5 upregulation by T cells[7].

Verified Bioactivity

1.The ED50 is <70 ng/mL, measured in a neutralization assay using TF-1 cells in the presence of 0.5 ng/mL IL-4.
2.Measured by its ability to inhibit IL-4-dependent proliferation of TF-1 human erythroleukemic cells. The ED50 for this effect is 3-15 ng/mL in the presence of 0.2 ng/mL IL-4.

MCE Validation Data

  • Purity - SDS-PAGE

    Purity - SDS-PAGE

    ≥ 95%, as determined by reducing SDS-PAGE.

  • Bioactivity - Cell-Based Assay

    Bioactivity - Cell-Based Assay

    Measured by its ability to inhibit IL-4-dependent proliferation of TF-1 human erythroleukemic cells.The ED50 for this effect is 3.143 ng/mL in the presence of 0.2 ng/mL IL4, corresponding to a specific activity is3.18×105 units/mg.

Technical Parameters

  • Species Human
  • Source HEK293
  • Tag Tag Free
  • Accession

    P24394-1 (G24-H232)

  • Gene ID
    3566  [NCBI]
  • Molecular Construction
    • N-term
    • IL-4Rα (G24-H232)
      Accession # P24394-1
    • C-term
  • Protein Length

    Extracellular Domain

  • Synonyms

    IL4R; IL4RA; Interleukin 4 Receptor; Interleukin-4 Receptor Alpha Chain; Interleukin-4 Receptor Subunit Alpha; IL-4R Subunit Alpha; CD124; IL4R Nirs Variant 1; IL-4 Receptor Subunit Alpha; CD124 Antigen; Interleukin 13 Receptor; IL-4R-Alpha; IL-4RA

  • AA Sequence

    GNMKVLQEPTCVSDYMSISTCEWKMNGPTNCSTELRLLYQLVFLLSEAHTCIPENNGGAGCVCHLLMDDVVSADNYTLDLWAGQQLLWKGSFKPSEHVKPRAPGNLTVHTNVSDTLLLTWSNPYPPDNYLYNHLTYAVNIWSENDPADFRIYNVTYLEPSLRIAASTLKSGISYRARVRAWAQCYNTTWSEWSPSTKWHNSYREPFEQH

  • Molecular Weight

    Approximately 37-55 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.

  • Glycosylation

    Yes

  • Purity

    ≥ 95%, as determined by reducing SDS-PAGE.

Product Properties

Appearance

Lyophilized powder.

Formulation

1.Lyophilized from a 0.22 μm filtered solution of PBS.
2.Lyophilized from a 0.22 μm filtered solution of 20 mM PB, 150 mM NaCl, pH 7.4.
Please refer to the lot-specific COA for specific buffer information.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Storage & Stability

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Shipping

Room temperature in continental US; may vary elsewhere.

References

[1]. Keegan AD,et al. An IL-4 receptor region containing an insulin receptor motif is important for IL-4-mediated IRS-1 phosphorylation and cell growth. Cell. 1994 Mar 11;76(5):811-20. [Content Brief]

[2]. Zurawski SM, et al. The primary binding subunit of the human interleukin-4 receptor is also a component of the interleukin-13 receptor. J Biol Chem. 1995 Jun 9;270(23):13869-78. [Content Brief]

[3]. Rolling C, et al. IL4 and IL13 receptors share the gamma c chain and activate STAT6, STAT3 and STAT5 proteins in normal human B cells. FEBS Lett. 1996 Sep 9;393(1):53-6. [Content Brief]

[4]. Jung T, et al. Soluble human interleukin-4 receptor is produced by activated T cells under the control of metalloproteinases. Int Arch Allergy Immunol. 1999 May;119(1):23-30. [Content Brief]

[5]. HageT,etal.Crystalstructureoftheinterleukin-4/receptoralphachaincomplexrevealsamosaicbindinginterface.Cell.1999Apr16;97(2):271-81. [Content Brief]

[6]. Kashiwada M, et al. Immunoreceptor tyrosine-based inhibitory motif of the IL-4 receptor associates with SH2-containing phosphatases and regulates IL-4-induced proliferation. J Immunol. 2001 Dec 1;167(11):6382-7. [Content Brief]

[7]. Myburgh E, et al. Murine IL-4 is able to signal via chimeric human IL-4Ralpha/mouse gamma-chain receptor. Mol Immunol. 2008 Mar;45(5):1327-36. [Content Brief]

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Calculators

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The Specific Activity Calculator Equation
  • Specific Activity (Unit/mg)
  • Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) Specific Activity (Unit/mg)
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100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     ≥ 95%, as determined by reducing SDS-PAGE.

  • 1/16

    MedChemExpress Validation
     Measured by its ability to inhibit IL-4-dependent proliferation of TF-1 human erythroleukemic cells.The ED50 for this effect is 3.143 ng/mL in the presence of 0.2 ng/mL IL4, corresponding to a specific activity is3.18×105 units/mg.

  • IL-4R alpha/CD124 Protein, Human (HEK293) Product Image
    1/16
  • Experimental Validation Results for IL-4R alpha/CD124 Protein, Human (HEK293)
  • Experimental Validation Results for IL-4R alpha/CD124 Protein, Human (HEK293)
  • IL-4R alpha/CD124 Protein, Human (HEK293) Product Image