Human Interferon-beta 1b Recombinant

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Human Interferon-beta 1b Recombinant

$70.00$160.00


accession P01574


Source Optimized DNA sequence encoding Human beta Interferon 1b mature chain was expressed in Chinese Hamster Ovary cells.
Molecular weight Native Human Interferon-beta, generated by the proteolytic removal of the signal peptide and propeptide,the molecule has a calculated molecular mass of approximately 23 kDa. Recombinant Interferon-beta 1b is a monomeric protein consisting of 166 amino acid residue subunits, and migrates due to glycosylation as an approximately 23 kDa protein under non-reducing conditions and reducing conditions in SDS-PAGE.
Purity >95%, as determined by SDS-PAGE and HPLC
Biological Activity Theactivity was determined by the cytopathic inhibition assay of human WISH cells infected with the ECMV virus, and determined to bex107 IU/mg.
Protein Sequence MTNKCLLQIA LLLCFSTTAL SMSYNLLGFL QRSSNFQCQK LLWQLNGRLE YCLKDRMNFD IPEEIKQLQQ FQKEDAALTI YEMLQNIFAI FRQDSSSTGW NETIVENLLA NVYHQINHLK TVLEEKLEKE DFTRGKLMSS LHLKRYYGRI LHYLKAKEYS HCAWTIVRVE ILRNFYFINR LTGYLRN
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 Interferon betabwas lyophilized from a.2 μm filtered solution containingmM NaOAc pH.5..
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 P17181
Interactor P48551 INAR2_HUMAN
Biological Process Antiviral-defense
Molecular function Cytokine

Methods

Islet isolation, culture, and exposure to different beta-cell inhibitory substances

  • Following the initial culture period, islets were cultured for an additional 6–24 hours in CMRL 1066 containing antibiotics, 2 mM glutamine and one of the following supplements: 0.5 mM sodium palmitate solubilized in 0.5% (weight/volume) fatty…
  • Following the initial culture period, islets were cultured for an additional 6–24 hours in CMRL 1066 containing antibiotics, 2 mM glutamine and one of the following supplements: 0.5 mM sodium palmitate solubilized in 0.5% (weight/volume) fatty acid and lipopolysaccaride free bovine serum albumin (BSA) ; recombinant human Interleukin 1beta (IL-1ß (50 units/ml) and Interferon-gamma (IFN-γ) (1,000 units/ml) ; 100 mM hydrogen peroxide; 2 mM DETA/NO or 10 mM streptozotocin (STZ) .
  • To some of the groups 10 µM of Imatinib was added at different time points prior to the addition of test substances given above.
  • To controls equal amounts of vehicle (DMSO) were supplemented.

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Western blotting for NF-κB, IκB-α and Smad7

  • Interferon gamma (IFN-γ) 50 μl (100 ng/ml) was added to each dish in the experimental studies.
  • The cytoplasmic and nuclear extracts were washed with ice-cold PBS and lysed in a 0.5 ml/well lysis buffer (150 mmol/l NaCl,…
  • Interferon gamma (IFN-γ) 50 μl (100 ng/ml) was added to each dish in the experimental studies.
  • The cytoplasmic and nuclear extracts were washed with ice-cold PBS and lysed in a 0.5 ml/well lysis buffer (150 mmol/l NaCl, 20 mmol/l Tris, pH 7.5, 0.1% Triton X-100, 1 mmol/l phenylmethylsulfonyl fluoride [PMSF] and 10 μg/ml aprotonin) as modified from the reports of Kim et al.
  • and Moon et al.
  • [

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Replicative control of HCV in JFH-1/Huh7.5.1 system with conditioned media (CM) from pU/UC-transfected pDC-GEN2.2 cells.

Recombinant Interferon proteins
  • E) Recombinant Type III Interferons in the absence of CM at the same concentrations as found in the CM (IL-28A/IFNλ2: 1500 pg/mL; IL-28B/IFNλ3: 10 pg/mL; IL-29/IFNλ1: 500 pg/mL) were added to JFH-1 infected Huh7.5.1 cells.

CIK induction and intravenous infusion

  • Venous blood was collected prior to chemotherapy or at least one month following chemotherapy.
  • For the offspring (HLA haploidentical donors), the routine blood tests, and liver and kidney function tests were normal and negative for hepatitis A virus-IgM,…
  • Venous blood was collected prior to chemotherapy or at least one month following chemotherapy.
  • For the offspring (HLA haploidentical donors), the routine blood tests, and liver and kidney function tests were normal and negative for hepatitis A virus-IgM, hepatitis B surface antigen antibody , hepatitis B e --hepatitis B core-hepatitis C virus (HCV)-HCV-IgG, Syphilis-and human immunodeficiency virus-.
  • As reported in our previous study (2 atmosphere.
  • On the day of culture, 1,000 U/ml human recombinant interferon-γ , 500 U/ml recombinant human interleukin (rhIL)-1α and 1,000 U/ml rhIL-2 (Quangang Pharmaceutical Co., , , ) were added.
  • Four days later, 1,000 U/ml rhIL-2 was added and the cells were transferred into a GT-T610 culture bag .
  • Cell growth was observed every other day and cells were stained with 0.4% trypan blue and the viable cells were counted.
  • Following 18 days of culture, cells were infused once daily (>1×109 cells; viability rate, >95%).
  • A cycle of treatment comprised of between three and five infusions and >5×109 cells were infused per cycle.
  • Prior to infusion, the surface immunophenotype of CIKs was determined by a standard fluorescence cytometry labeling protocol using fluorochrome-conjugated monoclonal mouse anti-human antibodies against CD3, CD8, CD4, CD45, CD19 and CD16CD56 .
  • Assays for the detection of bacteria, mildew and endotoxin were performed.
  • Briefly, the presence of bacteria and mildew was investigated by incubating cultured CIKs on agar at 37°C, with subsequent inspection for the growth of microbial contaminants.
  • Endotoxin levels were determined using a chromogenic endotoxin assay kit ( Crab Reagent Manufactory, , , , ), according to the manufacturer’s instructions.
  • Only sterile and endotoxin-free preparations were used clinically.

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(A) Nitrite production by HT-29 cells following 48 h of treatment with cytokines.

100 U/ml interferon
  • Effect of 0–50 ng/ml TNF-α on nitrite production induced by 100 U/ml IFN-γ and 10 ng/ml IL-1α in HT-29 cells following 48 h of treatment.

Invasion assay

  • Cell migration through Matrigel-coated filters was measured using Transwell chambers with 8 μm-pore polycarbonate filters coated with Matrigel matrix as previously described (4 cells/well in the upper compartment of each invasion chamber and incubated for…
  • Cell migration through Matrigel-coated filters was measured using Transwell chambers with 8 μm-pore polycarbonate filters coated with Matrigel matrix as previously described (4 cells/well in the upper compartment of each invasion chamber and incubated for 24 h in the absence or presence of 100 U/ml interferon (IFN)-γ , 10 ng/ml interleukin (IL)1-α or 25 ng/ml tumor necrosis factor (TNF)-α , plus extract of Cnidiihizoma (0.1–5 mg/ml) or 1400W (0.5 mM).
  • Non-migrating cells on the upper surface of the membrane were gently scrubbed with a cotton swab, and the invading cells on the lower surface were fixed with 100% methanol and stained with hematoxylin and eosin Y solution (RICCA Chemical Company, Charlotte, NC, USA).
  • The number of cells was counted under a microscope at a magnification of ×100.

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