Human IL-35 (IL12A & EBI3 Heterodimer) HEK293 Overexpression Lysate 产品信息
Product Description
This Human IL-35 (IL12A & EBI3 Heterodimer) overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IL-35 (IL12A & EBI3 Heterodimer) protein (Cat: CT065-H2508H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human IL27B (NP_005746.2)(Met1-Lys229) was fused with a flag tag at the C-terminus, constructed the plasmid 1; A DNA sequence encoding the human IL12A (NP_000873.2)(Met1-Ser219) was fused with a polyhistidine tag at the C-terminus, constructed the plasmid 2. The two plasmids were co-expressed and the human IL27B&IL12A heterodimer was purified.
Molecule Mass
The recombinant heterodimer of human IL27B&IL12A comprises 425 (217+208) amino acids and has a calculated molecular mass of 48.3(24.3+24) KDa.
Human IL-35 (IL12A & EBI3 Heterodimer) 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 IL-35 (IL12A & EBI3 Heterodimer) HEK293 Overexpression Lysate Alternative Names
Human CLMF Overexpression Lysate;Human IL-12A Overexpression Lysate;Human NFSK Overexpression Lysate;Human NKSF1 Overexpression Lysate;Human P35 Overexpression Lysate
IL-35 (IL12A & EBI3 Heterodimer) Background Information
IL12A is a subunit of a cytokine that acts on T and natural killer cells, and has a broad array of biological activities. IL12A, together with IL27B, form a disulfide-linked heterodimer: IL12A&IL27B. IL12A&IL27B is required for the T-cell-independent induction of IFN-gamma, and is important for the differentiation of both Th1 and Th2 cells. The responses of lymphocytes to this cytokine are mediated by the activator of transcription protein STAT4. Nitric oxide synthase 2A (NOS2A/NOS2) is found to be required for the signaling process of IL12A&IL27B in innate immunity.
Full Name
interleukin 12A
References
Wolf S.F., et al.,(1991), Cloning of cDNA for natural killer cell stimulatory factor, a heterodimeric cytokine with multiple biologic effects on T and natural killer cells. J. Immunol. 146:3074-3081. Gubler U., et al., (1991), Coexpression of two distinct genes is required to generate secreted bioactive cytotoxic lymphocyte maturation factor.Proc. Natl. Acad. Sci. U.S.A. 88:4143-4147. Batten M., et al.,(2007), The biology and therapeutic potential of interleukin 27.J. Mol. Med. 85:661-672.
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