Human Interferon alpha-B HEK293 Overexpression Lysate: Product Information
This Human Interferon alpha-B overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Interferon alpha-B protein (Cat: 10347-HNCH) from the overexpression lysate was verified.
A DNA sequence encoding the human IFNA8 (NP_002161.2)(Cys24-Glu189) was expressed and purified with two additional amino acids (Gly & Pro ) at the N-terminus.
The secreted recombinant human IFNA8 consists of 168 amino acids and predicts a molecular mass of 19.6 KDa. The apparent molecular mass of the protein is approximately 26 KDa in SDS-PAGE under reducing conditions due to glycosylation.
Human Interferon alpha-B HEK293 Overexpression Lysate: Usage Guide
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.
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.
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human Interferon alpha-B HEK293 Overexpression Lysate: Alternative Names
Human IFN-alphaB Overexpression Lysate
Interferon alpha-B Background Information
Interferon alpha-B, also known as IFNA8, belongs to thealpha/beta interferon family. Interferons areproteinsmade and released byhost cellsin response to the presence ofpathogenssuch asviruses,bacteria,parasitesortumorcells. Interferon stimulates the production of two enzymes: a protein kinase and an oligoadenylate synthetase. They also allow for communication between cells to trigger the protective defenses of theimmune systemthat eradicate pathogens or tumors.Interferons also activate immune cells, such asnatural killer cellsandmacrophages. They increase recognition of infection or tumor cells by up-regulatingantigen presentation toT lymphocytes. They also increase the ability of uninfected host cells to resist new infection by virus. Certain symptoms, such as aching muscles and fever, are related to the production of IFNs during infection. Produced by macrophages, IFN-alpha have antiviral activities.
interferon, alpha 8
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Kempaiah P. et al., 2012, Hum Genet. 131 (8): 1375-91.
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