Human Interferon-alpha 2a Recombinant

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Human Interferon-alpha 2a Recombinant

$70.00$880.00


accession P01563


Source Optimized DNA sequence encoding Human Interferon-alpha mature chain was expressed in Escherichia Coli.
Molecular weight Recombinant human IFN-alphaa, generated by the proteolytic removal of the signal peptide and propeptide, and has a calculated molecular mass of approximately kDa. Recombinant Interferonalphaais a disulfide-linked monomeric protein consisting of amino acid residue subunits, and migrates as an approximately19 kDa protein under non-reducing conditions and reducing conditions in SDS-PAGE.
Purity >98%, as determined by SDS-PAGE and HPLC
Biological Activity Theactivity was determined bya viral resistance assay of Human WISH cells, andwas found to be in the range ofx108 IU/mg.
Protein Sequence MALTFALLVA LLVLSCKSSC SVGCDLPQTH SLGSRRTLML LAQMRKISLF SCLKDRHDFG FPQEEFGNQF QKAETIPVLH EMIQQIFNLF STKDSSAAWD ETLLDKFYTE LYQQLNDLEA CVIQGVGVTE TPLMKEDSIL AVRKYFQRIT LYLKEKKYSP CAWEVVRAEI MRSFSLSTNL QESLRSKE
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 Interferon alphaawas lyophilized from a.2 μm filtered PBS solution pH.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 P17181
Interactor P48551 INAR2_HUMAN
Biological Process Antiviral-defense
Molecular function Cytokine

Methods

Cytokine treatment

  • After 2 passages, cells were seeded into 6-well plates .
  • At confluence, cytokine treatment was performed during 48 hours using human recombinant TNF-α 100 U/ml or IFN-γ 200 U/ml in serum-free media, otherwise as above.
  • Experiments were ended by removal and freezing of the supernatants and addition of lysis buffer to the cell monolayer, see below.

V protein expression counteracts the HCV-inhibitory effects of added type I and type III interferons in HFLC.

IFN-β
  • Cells were washed three times then refed with HDM containing serial 10-fold dilutions of IFN-β, IL-28A, or IL-29.

IFN-γ treatment induced the deposition of active histone modifications throughout the HLA-DRA gene.

500 U/ml IFN-γ
  • Transcript levels for A431 cells are shown before and after treatment with 500 U/ml of IFN-γ for 24 hrs.

Decrease of ISG15 conjugates by reducing agents.

human IFN-β
  • 24 h post-transfection, the cells were induced with IFN-β (1,000 units/ml).

Cell Culture and Transfection

  • HeLa B cells (ECACC # 85060701) were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% heat-inactivated calf serum (Sigma-Aldrich), penicillin (100 units/ml) (Gibco), streptomycin (100 µg/ml) and Non-Essential Amino Acids Solution 10 mM (100x) (Gibco) at 37°C with 5% CO2 in a humidified incubator.
  • Cells were transiently transfected with GeneJuice according to the manufacturer’s instructions.
  • For co-transfections, the total plasmid amount for each transfection was normalized by the addition of appropriate amount of empty vectors.
  • For cytokine stimulation, 1,000 units/ml of human IFN-β was used.