Human IL-27 CHO Stable Overexpression Lysate: Product Information
This Human IL-27 overexpression lysate was created in CHO Stable Cells and intented for use as a Western blot (WB) positive control. Purification of IL-27 protein (Cat: 10076-H08S) from the overexpression lysate was verified.
CHO Stable Cells
A DNA sequence encoding the IL27 complex composed of IL27B subunit (NP_005746.2) (Met 1-Lys 229) and the N-terminally truncated IL27p28 (IL27A) subunit (NP_663634.2) (Phe 29-Pro 243) linked by an polypeptide linker was fused with a polyhistidine tag at the C-terminus.
The recombinant human IL27 complex (IL27B & IL27A) consists of 450 amino acids after removal of the signal peptide and has a calculated molecular mass of 56 KDa as estimated in SDS-PAGE under reducing conditions.
Human IL-27 CHO Stable 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 IL-27 CHO Stable Overexpression Lysate: Alternative Names
Human IL-27 Overexpression Lysate; Human IL-27A Overexpression Lysate; Human IL27A Overexpression Lysate; Human IL27p28 Overexpression Lysate; Human IL30 Overexpression Lysate; Human p28 Overexpression Lysate
IL-27 Background Information
IL-27 protein is a member of the IL-6 superfamily, which is expressed on monocytes, endothelial cells and dendritic cells. IL-27 protein is also referred as the IL-12 p35-related protein, p28, is one subunit of a heterodimeric cytokine complex, and associates with another subunit EBI3 (EBV-induced gene 3), an IL-12 p4-related protein (IL-27B). IL-27 protein is an early product of activated antigen-presenting cells and drives rapid clonal expansion of naive CD4(+) T cells and plays a role in the early regulation of Th1 cells initiation which drives efficient adaptive immune response. IL-27 protein has an antiproliferative activity on melanomas through WSX-1/STAT1 signaling. Thus, IL-27 protein may be an attractive candidate as an antitumor agent applicable to cancer immunotherapy.
Hisada M, et al. (2004) Potent antitumor activity of interleukin-27. Cancer Res. 64(3): 1152-6.
Larousserie F, et al. (2005) Analysis of interleukin-27 (EBI3/p28) expression in Epstein-Barr virus- and human T-cell leukemia virus type 1-associated lymphomas: heterogeneous expression of EBI3 subunit by tumoral cells. Am J Pathol. 166(4): 1217-28.
Seita J, et al. (2008) Interleukin-27 directly induces differentiation in hematopoietic stem cells. Blood. 111(4): 1903-12.
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