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Your Position: Home > Cytokines > IL23A & IL12B > ILB-H52W5

Human IL-23 alpha&IL-12 beta Heterodimer Protein, His Tag&Tag Free

INTRODUCTION

QC & Assay Protocol

Citation & Background

Synonym
IL-23 alpha & IL-12 beta
Source
Human IL-23A&IL-12B Heterodimer Protein, His Tag&Tag Free (ILB-H52W5) is expressed from human 293 cells (HEK293). It contains AA Arg 20 - Pro 189 (IL23A) & Ile 23 - Ser 328 (IL12B) (Accession # Q9NPF7-1 (IL23A) & P29460-1 (IL12B)).
Predicted N-terminus: His (IL23A) & Ile 23 (IL12B)
Request for sequence
Molecular Characterization
Human IL-23A&IL-12B Heterodimer Protein, His Tag&Tag Free, produced by co-expression of IL-23 alpha and IL-12 beta, has a calculated MW of 19.5 kDa (IL-23 alpha) and 34.7 kDa (IL-12 beta). Subunit IL-23 alpha is fused with a polyhistidine tag at the N-terminus and subunit IL-12 beta contains no tag. The reducing (R) protein migrates as 20 kDa (IL-23 alpha) and 37-45 kDa (IL-12 beta) respectively due to glycosylation.
Endotoxin
Less than 1.0 EU per μg by the LAL method.
Purity
>95% as determined by SDS-PAGE.

Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
1. -20°C to -70°C for 12 months in lyophilized state;
2. -70^°C for 3 months under sterile conditions after reconstitution.

SDS-PAGE

Human IL-23A&IL-12B Heterodimer Protein, His Tag&Tag Free on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.

Bioactivity-ELISA

Human IL-23A&IL-12B Heterodimer Protein, His Tag&Tag FreeHuman IL-23A&IL-12B Heterodimer Protein, His Tag&Tag Free (Cat. No. ILB-H52W5) ELISA bioactivity

Immobilized Human IL-23A&IL-12B Heterodimer Protein, His Tag&Tag Free (Cat. No. ILB-H52W5) at 2 μg/mL (100 μL/well) can bind Anti IL23A&IL12B P40 domain, Human IgG1 with a linear range of 0.05-3 ng/mL (QC tested).

Protocol

Immobilized Human IL-23 R, Fc Tag (Cat. No. ILR-H5254) at 10 μg/mL (100 μL/well) can bind Human IL-23A&IL-12B Heterodimer Protein, His Tag&Tag Free (Cat. No. ILB-H52W5) with a linear range of 20-78 ng/mL (Routinely tested).

Protocol

Bioactivity-SPR

Captured Human IL-23 R, Fc Tag (Cat. No. ILR-H5254) on CM5 chip via anti-human IgG Fc antibodies surface can bind Human IL-23A&IL-12B Heterodimer Protein, His Tag&Tag Free (Cat. No. ILB-H52W5) with an affinity constant of 5.36 nM as determined in a SPR assay (Biacore T200) (Routinely tested).

Protocol

Bioactivity-Cell based assay

Human FcRn Heterodimer Protein Cell_Base

Human IL-23A&IL-12B Heterodimer Protein, His Tag&Tag Free (Cat. No. ILB-H52W5) stimulates secretion of IL-17 by mouse spleen cells. The ED50 for this effect is 0.1055-0.1635 ng/mL (Routinely tested).

Protocol

Cell based assay shows that the secretion of IL-17 induced by Human IL-23A&IL-12B Heterodimer Protein, His Tag&Tag Free (Cat. No. ILB-H52W5) is inhibited by increasing concentration of the anti-human IL-23 neutralizing antibody. The IC50 is between 0.67-1.50 ng/mL (Routinely tested).

Protocol

Background
Interleukin-23 subunit alpha (IL-23 alpha) can associates with IL12B to form the IL-23 interleukin, a heterodimeric cytokine which functions in innate and adaptive immunity. IL-23 may constitute with IL-17 an acute response to infection in peripheral tissues. IL-23 binds to a heterodimeric receptor complex composed of IL12RB1 and IL23R, activates the Jak-Stat signaling cascade, stimulates memory rather than naive T-cells and promotes production of proinflammatory cytokines. IL-23 induces autoimmune inflammation and thus may be responsible for autoimmune inflammatory diseases and may be important for tumorigenesis.
References
- (1) Oppmann B., et al., 2000, Immunity 13:715-725.
- (2) Parham C., et al., 2002, J. Immunol. 168:5699-5708.
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.