Active Recombinant Human Iinterleukin 23, Alpha Subunit P19

• Cat.No.: L23A-269H
• Product Overview: Recombinant Human IL23A is expressed inHuman 293 cellsas a heterodimeric glycoprotein composed of two disulfide-linked subunits (p40 cystine linked to p19).

Specification

• Species: Human
• Source: HEK293
• Tag: Non
• Description: Production in Human 293 cells offers authentic glycosylation. Cytokines produced in Sf9 or Sf21 cells have posttranslational modifications which are not humanlike. This gene encodes a subunit of the heterodimeric cytokine interleukin 23 (IL23). IL23 is composed of this protein and the p40 subunit of interleukin 12 (IL12B). The receptor of IL23 is formed by the beta 1 subunit of IL12 (IL12RB1) and an IL23 specific subunit, IL23R. Both IL23 and IL12 can activate the transcription activator STAT4, and stimulate the production of interferon-gamma (IFNG). In contrast to IL12, which acts mainly on naive CD4 (+) T cells, IL23 preferentially acts on memory CD4 (+) T cells.
• Endotoxin: Endotoxin-free.
• Purity: >95%. The protein was resolved by SDS polyacrylamide gel electrophoresis and the gelwas stained with Coomassie blue.
• Activity: The specific activity was determined by the dose-dependent secretion of IL-17 from mouse splenocytes activated with 10ng/mL PMA.
• Solubility: It is recommended to reconstitute the lyophilized Bone Morphogenetic Protein-6 in sterile 20mM AcOH (acetic Acid) not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
• Storage: -20℃.

Gene Information

• Gene Name: IL23A interleukin 23, alpha subunit p19 [Homo sapiens]
• Synonyms: P19; SGRF; IL-23; IL-23A; IL23P19; MGC79388; IL23A ;interleukin-23 subunit alpha ; IL-23-A ; IL-23p19 ; IL-23 subunit alpha ; interleukin 23 p19 subunit ; interleukin-23 subunit p19 ; JKA3 induced upon T-cell activation
• Gene ID: 51561
• mRNA Refseq: NM_ 016584
• Protein Refseq: NP_057668
• MIM: 605580
• UniProt ID: Q9NPF7
• Chromosome Location: 12q13.3
• Pathway: Cytokine-cytokine receptor interaction; Jak-STAT signaling pathway
• Function: cytokine activity; protein binding

Citation

A Role for RUNX3 in Inflammation-Induced Expression of IL23A in Gastric Epithelial Cells

Journal: Cell reports    PubMed ID: 25008775    Data: 2015/1/20

Authors: Yit Teng Hor, Dominic Chih-Cheng Voon, Yoshiaki Ito

Article Snippet:For measurement of IL-23 in the supernatants harvested from cultured cells, ELISA was performed using the Human IL-23 Ready-SET-Go! kit (eBioscience) and the Human IL23A/p19 ELISA kit (Creative BioMart) as per the manufacturer’s instructions.. To induce the secretion of IL-23, gastric epithelial cell lines and the monocytic cell line THP-1 were stimulated with TNF-α, H. pylori , or LPS for 24 hr.To induce the secretion of IL-23, gastric epithelial cell lines and the monocytic cell line THP-1 were stimulated with TNF-α, H. pylori , or LPS for 24 hr.

(A) IL23A mRNA expression was induced by exogenous RUNX3 in multiple RUNX3-negative gastric cancer cell lines. GFP-positive transfected cells were enriched by FACS at 24 hr and 48 hr posttransfection and analyzed by qRT-PCR. Normalized IL23A levels are expressed relative to untransfected control values. (B) RUNX3 specifically induced IL23A in gastric epithelial cells. AGS cells transduced with the indicated viruses were analyzed by qRT-PCR for the expression of the IL-12 family of cytokine genes. Normalized data are presented relative to the Lenti-control sample (mean ± SEM; n = 3) (u.d., undetected). (C) RUNX3 mediates its effect through the proximal RUNX sites B, C, and D of the IL23A promoter. Mutation and deletion variants of the IL23A-1200 reporter construct were transiently transfected into KATOIII cells together with either control or RUNX3 expression vectors. Normalized luciferase activities are expressed relative to the values of control samples for each construct. Data presented are derived from independent biological triplicates (mean ± SEM). (D) Physical occupancy of RUNX3 on the IL23A promoter in Lenti-RUNX3-transduced AGS cells was detected by ChIP analysis using polyclonal or monoclonal RUNX3-specific antibodies. Enrichment of the indicated genomic fragments was detected by qPCR and expressed relative to input DNA. Nonspecific immunoglobulin G from rabbit (rIgG) and mouse (mIgG) served as negative controls; H3K9Ace antibody was used as a positive control. NS, nonspecific region; pAb, polyclonal antibody; mAbs, monoclonal antibodies. *p < 0.05; **p < 0.01. See also .

(A and B) The effects of various proinflammatory stimuli on the expression of IL23A . AGS cells were treated with the indicated (A) cytokines and (B) ligands and agonists of various TLRs and NLRs for 6 hr prior to qRT-PCR measurement of IL23A transcript. Normalized IL23A mRNA levels are expressed relative to that of untreated controls (Mock). Data presented are derived from biological triplicates (mean ± SEM). (C) Cooperative induction of IL23A by RUNX3 and TNF-α. AGS cells were transduced with the indicated lentiviruses for 48 hr and treated with TNF-α for 6 hr for qRT-PCR analysis (top) or 18 hr for western blot analysis (bottom). Top: normalized IL23A mRNA levels are plotted relative to the basal values of Lenti-control-infected cells (mean ± SEM; n = 3). Bottom: whole-cell lysates were analyzed for IL23A, RUNX3, and α-tubulin (loading control) expression by western blotting. (D and E) Sequence requirement for TNFα induction of IL23A promoter. Wild-type and mutant variants of the IL23A promoter reporter constructs were transiently transfected into the cells together with either control or RUNX3 expression vector for 24 hr followed by TNF-α treatment for 24 hr. Normalized luciferase activities are presented relative to those of untreated control samples (mean ± SEM; n = 3). *p < 0.05; **p < 0.01. See also .

(A) H. pylori activates IL23A in a dose-dependent manner. AGS cells were cocultured with different MOIs of live or heat-killed H. pylori for 6 hr prior to qRT-PCR quantification of IL23A transcript. The normalized values are presented relative to those of uninfected control samples (MOI 0) (mean ± SEM; n = 3). (B) IL23A induction by H. pylori requires activation of the SHP-2/ERK pathway by oncoprotein CagA. AGS cells were preincubated with 50 μM of SHP-2 inhibitor (NSC87877) for 3 hr followed by infection with wild-type or ΔCagA strains of H. pylori at MOI100 or vehicle (Mock) for 18 hr. The normalized values are presented relative to those of uninfected control samples (mean ± SEM; n = 3). (C) TNF-α and H. pylori activate IL23A in a diverse range of gastric epithelial cell lines. Transformed and untransformed gastric epithelial cell lines were stimulated with TNF-α (10 ng/ml) or wild-type H. pylori (MOI100) for 6 hr. IL23A mRNA levels were determined by qRT-PCR and normalized values are presented relative to values of untreated controls of each cell line (mean ± SEM; n = 3). (D) The expression of IL23A in response to RUNX3, TNF-α, and H. pylori . AGS cells were transduced with the indicated lentiviruses for 48 hr prior to treatment with TNF-α and/or H. pylori for 6 hr for qRT-PCR analysis (top) or 18 hr for western blot analysis (bottom). Top: normalized IL23A levels are presented relative to those of untreated AGS cells infected with Lenti-control virus (mean ± SEM; n = 3). Bottom: whole-cell lysates were analyzed for IL23A, RUNX3 and α-tubulin protein expression by western blotting. (E) Both RUNX1 and RUNX3 transactivate the IL23A promoter in gastric epithelial cells. KATOIII and SNU16 were transiently transfected with the IL23A-1200 reporter construct together with the indicated expression vectors for 48 hr. Normalized reporter activities of each construct are plotted relative to the values of corresponding control samples. (F) The effects of RNAi knockdown of RUNX3 and RUNX1 on IL23A expression. HFE-145 cells were transfected with control small interfering RNA (siRNA) (siCtrl), RUNX3 siRNA (siRX3), and/or RUNX1 siRNA (siRX1) for 48 hr prior to treatment with H. pylori and TNF-α for 6 hr and qRT-PCR measurements. Normalized IL23A levels are plotted relative to those of untreated siCtrl sample (mean ± SEM; n = 3). *p < 0.05; **p < 0.01. See also .

Q&As

Is this bioactive cyno IL-23 heterodimer (p19/p40) or just the cyno IL-23p19 subunit?

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