Human IFN-omega HEK293 Overexpression Lysate 产品信息
Product Description
This Human IFN-omega overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IFN-omega protein (Cat: 10353-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human IFNω (NP_002168.1) (Cys 24-Ser 195) was fused with the Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human IFNω/Fc chimera is a disulfide-linked homodimer. The reduced monomer comprises 409 amino acids with a predicted molecular mass of 46.6 kDa. As a result of glycosylation, rh IFNω/Fc monomer migrates as an approximately 50 kDa band in SDS-PAGE under reducing conditions.
Human IFN-omega HEK293 Overexpression Lysate Usage Guide
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Human IFN-omega HEK293 Overexpression Lysate Alternative Names
Human IFN-omega 1 Overexpression Lysate;Human IFNW1 Overexpression Lysate;Human interferon omega-1 Overexpression Lysate
IFN-omega Background Information
IFNs are a large family of proteins having antiviral, antiproliferative, and immunomodulatory effects, and are divided into two major classes, type I and type I I, on the basis of differences in receptor binding and nucleotide sequence. Type I IFNs consist of IFN α, β, τ, and ω and bind to the type I IFN receptor, whereas IFN-γ is the only type I I IFN and is specific for the type I I IFN receptor. Human IFN-ω, was identified by three independent groups in 1985 and is structurally related to IFN-α and -β. Both human IFN-ω and IFN-α are produced by virally induced leukocytes and have similar antiviral activities on human cell lines, and a sizeable proportion (at least 1%) of the total antiviral activity of leukocyte IFN is contributed by IFN-ωl. In addition, it was reported that IFN-ω could inhibit the growth of human tumors in vivo.
Full Name
interferon, omega 1
References
Capon D.J., et al.,(1985), Two distinct families of human and bovine interferon-alpha genes are coordinately expressed and encode functional polypeptides. Mol. Cell. Biol. 5:768-779. Adolf G.R., et al., (1991), Human interferon omega 1: isolation of the gene, expression in Chinese hamster ovary cells and characterization of the recombinant protein.Biochim. Biophys. Acta 1089:167-174. Humphray S.J., et al.,(2004), DNA sequence and analysis of human chromosome 9.Nature 429:369-374.
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