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Human Interleukin-10 Recombinant

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$70.00$4,700.00

SKU: RKP22301 Tags: , , , ,

Description

Accession
P22301
Source
Optimized DNA sequence encoding Human Interleukin-10 mature chain was expressed in Escherichia Coli.
Molecular weight
Native HumanInterleukin-10 is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately kDa. Recombinant Human Interleukin-10 is a monomer protein consisting of161 amino acid residue subunits, and migrates as an approximately19 kDa protein under non-reducing andreducing conditions in SDS-PAGE.
Purity
>95%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED50 determined by dose-dependent co-stimulation (with murine IL-4) of MC/9 cells. The ED50 was found to be ≤ ng/ml, corresponding to a specific activity of ≥x units/mg.
Protein Sequence
MHSSALLCCL VLLTGVRASP GQGTQSENSC THFPGNLPNM LRDLRDAFSR VKTFFQMKDQ LDNLLLKESL LEDFKGYLGC QALSEMIQFY LEEVMPQAEN QDPDIKAHVN SLGENLKTLR LRLRRCHRFL PCENKSKAVE QVKNAFNKLQ EKGIYKAMSE FDIFINYIEA YMTMKIRN
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Presentation
Recombinant Interleukin-10 was lyophilized from a.2 μm filtered PBS solution pH7.0.
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.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 at° -° 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
Interactor
Molecular function

Methods

Cardiac adherent proliferating cells reduce Coxsackievirus B3-induced HL-1 apoptosis in a nitric oxide- and interleukin-10-dependent manner and require interferon-γ priming.

  • Bar graphs represent DiO+ Annexin V+/7AAD- HL-1 cells in cultures of uninfected (open bars) or CVB3-infected (closed bars) HL-1 with or without untreated or L-NAME treated CAPs or CAPs in the presence or absence of 1 µg/ml of anti-human IL-10 antibody (ab), or 1 µg/ml of anti-murine IFN–γ ab; n = 4/group.

Phagocytosis assay

  • To assess a potential role for, or IL-10 in phagocytosis of apoptotic cells by influenza-infected macrophages, mock- or Udorn-infected Mph1 and were incubated with an anti-Mer blocking antibody (& ; 5 µg/ml), (esearch HP3652L; 4 µg/ml) or recombinant human IL-10 (20 ng/ml) for 30 minutes before addition of the apoptotic neutrophils.
  • The reagents were also present during the 1 h macrophage-neutrophil co-culture.

Dendritic Cells and T Cells Function Assay

  • Cytokines were quantified using specific ELISA kits, following the manufacturer’s instructions.
  • IL-2, IL-12, and IL-10 kits were from , λ 1 (IL-29) and λ 2 (IL-28A, cross-reacting with IL-28B) were from& .

Isolation of monocytes and cell culture

  • Monocytes were obtained from buffy coats from healthy blood donors.
  • Written informed consents were obtained from all donors in accordance with the ethical principles set out in the declaration of.
  • This study was approved by the Medical Committee of the Academic Medical Center and the of the in , .
  • Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats using Lymphoprep density gradient centrifugation.
  • Monocytes were isolated by adherence to plastic and cultured in Iscove's medium'>modified medium'>Dulbecco's medium (IMDM;, , ) supplemented with 10% [v/v] heat-inactivated human pooled serum (HPS), penicillin (100 U/ml, , ), streptomycin (100 µg/ml) and ciproxin (5 μg/ml, , ) for 5 days in the presence of different cytokines: IFN-α (250 U/ml, , , ), IFN-β (250 U/ml), IFN-γ (250 U/ml and 50 U/ml) in combination with TNF-α (12.5 ng/ml, , , ), IL-4 (50 ng/ml), IL-10 (50 ng/ml), GM-CSF…

A: Ex vivo IFNγ ELISpot responses in patients with acute DHF who developed shock and those who did not develop shock and also in healthy dengue seropositive donors and dengue seronegative donors.

  • B: Spontaneously released IL-10 levels (pg/ml) in PBMC ELISpot supernatants in patients with shock and those without shock The mean spontaneous release of IL-10 from PBMC in patients with shock was 19.65 (SD±21.28 pg/ml), and the mean levels in those who did not develop shock was 11.28 (SD±24.5) pg/ml (p = 0.08).

Characterization of a stable transfectant of PANC-1 cells expressing TGF-β1.

  • Mean concentration of VEGF, IL-10, the active form of TGF-β1 derived from PANC/Mock or PANC/TGF-β was analyzed by ELISA.

Ig Secretion by Human B Cells

  • Human B cells were isolated from buffy coats of healthy donors obtained from the New York Blood Center by negative selection with B cell isolation kit II .
  • The purity of the sorted B cells was more than 97%, as assessed by CD19 staining.
  • Purified B cells (5×104) were plated in a 96-well U-bottom plate in 200 µl of complete medium'>RPMI 1640 medium containing 10% FBS, 2mM glutamine, 100U/ml streptomycin, 100U/ml penicillin, 1mM sodium pyruvate, and 10mM HEPES (all from, , ).
  • The cells were treated with 20 µl of 293T cell supernatant transfected with Envwt or Env-fusion proteins in the presence of recombinant C40L (Enzo Life, , ) (200 ng/ml), interleukin-4 (IL-4) (10 ng/ml), and IL-10 (200 ng/ml) for 14 days.
  • The amount of Env or Env-fusion proteins was normalized based on an anti-Env ELISA using the 2G12 MAb (data not shown) and adjusted using mock…

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.

Mitogen stimulation

  • Differentiation of MBCs into ASCs in was initiated in bulk PBMC cultures based on a previously established protocol S.
  • aureus Cowan I protein A (SAC) and CpG.
  • IL-10 was added to the stimulation mix since a previous study showed that this enhanced the efficiency of MBC into ASC differentiation by more than 9 fold 6 cells/ml were added to 25 cm2 cell culture flasks .
  • Culture medium was supplemented with 50 ng/ml PWM lectin derived from Phytolacca americana , 1∶5000 Protein A from aureus, Cowan Strain , 2.5 µg/ml ODN 2006 ( nucleotide-human ) and 25 ng/ml recombinant human IL-10 and incubated at 37°C, 5% CO2 for 5 days.
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