Human Interleukin-17A Recombinant
Categories: IL-17 familyOther cytokines$70.00 – $2,700.00
Description
Accession
Q16552
Source
Optimized DNA sequence encoding Human Interleukin-17A mature chain was expressed in Escherichia Coli.
Molecular weight
Nativehuman Interleukin17A, generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately 16kDa. Recombinant IL-17A is a disulfide-linked homodimeric protein consisting of two 137 amino acid residue subunits,and migrates as an approximately 31kDa protein under non-reducing conditions and as a 16 kDa protein under reducing conditions in SDS-PAGE.
Purity
>95%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) was determined by the dose-dependent proliferation ofinduction of IL-6 in primary human foreskin fibroblasts was found to be approximately ng/ml., corresponding to a specific activity of > x units/mg
Protein Sequence
MTPGKTSLVS LLLLLSLEAI VKAGITIPRN PGCPNSEDKN FPRTVMVNLN IHNRNTNTNP KRSSDYYNRS TSPWNLHRNE DPERYPSVIW EAKCRHLGCI NADGNVDYHM NSVPIQQEIL VLRREPPHCP NSFRLEKILV SVGCTCVTPI VHHVA
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Presentation
Recombinant Human Interleukin-17 was lyophilized from a 0.2 μm filtered PBS solution pH7.5.
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers.
Storage
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at2° -8° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Interactor
Molecular function
Methods
Treg cells regulated Th17 response in HBV-LF .
- Compared to HC subjects (n = 30), Th17-associated cytokines such as IL-17, IL-21 and IL-22 significantly increased in serum of CHB patients (n = 77), however Treg-associated inhibitory cytokines IL-10 and TGF-β are similar in HC subjects and CHB patients.
Hepatic Stellate Cells Proliferation Assay
- HSCs were directly co-cultured with human CD4+, CD4+CD25+ and CD4+CD25− cells from age-and-sex matched HC and CHB subjects or incubated with human recombinant cytokine IL-17 at various concentrations (1, 3, 5 ng/mL) for 6 days.
- Cell proliferation of HSCs was monitored by a Cell-IQ system , which is a live cell imaging and analysis platform.
- Cell-IQ automatically recognizes cell type and records corresponding cell number every 15 minutes.
- Four high-power fields (hpf, 400×magnification) per well were chosen for consecutive cell monitoring and proliferation analysis.
- The cell proliferation rate was calculated using the following formula: (mean cell number in each hpf after incubation with stimulants)/(mean cell number in each hpf before incubation)×100%−1.
Regulation of membrane-bound and sCD127.
- PBMCs were cultured for 24 h in PMI 1640 supplemented with 5% FCS with the following stimuli: anti-C3/C28–coated beads (1:100 bead/cell ratio), IL-2 (100 U/mL ), IL-7 (10 ng/mL& ), interferon-γ (IFN-γ; 10 ng/mL ), IL-4 (10 ng/mL), IL-17 (10 ng/mL& ), TNF-α (10 ng/mL& ), IL-10 (10 ng/mL& ), IL-12 (10 ng/mL), rapamycin (sirolimus, 10 ng/mL), FK506 (tacrolimus, 10 ng/mL ), mycophenolate mofetil (MMF, 1 μg/mLoche), daclizumab (1 μg/mLoche), high glucose (33.3 mmol/L), and insulin (100 U/mL).
- PBMCs were subsequently stained with anti-CD4 Pacific blue (clone RPA-T4 ), anti-CD8 allophycocyanin-Cy7 (clone ), and anti-CD127 phycoerythrin (clone ), and surface expression of CD127 in gated CD4+ and CD8+ T cells was analyzed by flow cytometry.
- Supernatant from PBMC cultures was taken to measure sCD127 concentration.
Elevated frequencies of Th17 cells and reduced frequencies of Th1 cells in freshly isolated peripheral blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (BMMCs) from acute myeloid leukemia (AML) patients.
- (c) Total RNA was isolated from CD4+ T cells obtained from AML patients and HDs and reverse transcribed into cDNA and subsequently real time polymerase chain reaction for IL-17A and IFN-γ.