IL-35 Lysate | IL12A&IL27B lysate

Human IL-35 HEK293 Overexpression Lysate

• Applications: Western Blot
• Synonyms: IL-35; Human IL-35 HEK293 Overexpression Lysate; Human IL-35 (IL12A & EBI3 Heterodimer) HEK293 Cell Lysate (WB positive control); Human CLMF Overexpression Lysate; Human IL-12A Overexpression Lysate; Human IL-35 Overexpression Lysate; Human Interleukin-35 Overexpression Lysate; Human NFSK Overexpression Lysate; Human NKSF1 Overexpression Lysate; Human P35 Overexpression Lysate; interleukin 12A; IL12A&IL27B lysate

• Host: HEK293 Cells
• Form/Format: Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.; 1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).

• Applicable Applications for IL12A&IL27B lysate: Western Blot (WB)
• Species: Human
• Sequence Information: A DNA sequence encoding the p35 subunit of human IL12, termed as IL12A (P29459-1)(Met1-Ser219) was fused with the C-terminal DYKDDDDK-tagged Fc region of human IgG1 at the C-terminus, constructed the plasmid 1; A DNA sequence encoding the human IL27B (NP_005746.2)(Met1-Lys229) was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus, constructed the plasmid 2. The two plasmids were co-expressed and the heterodimer was purified.
• Preparation Method: Cell lysate was prepared by homogenization 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 (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
• 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 200uL pure water and boil for 2-5 min.; 3. Store the lyophilized cell lysate at 4 degree C. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80 degree C.
• Preparation and Storage: Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.

Related Product Information for IL12A&IL27B lysate

Description: This Human IL-35 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IL-35 protein from the overexpression lysate was verified.

Background: IL12A is a subunit of a cytokine that acts on T and natural killer cells and has a broad array of biological activities. The cytokine is a disulfide-linked heterodimer composed of the 35-kD subunit encoded by this gene, and a 40-kD subunit that is a member of the cytokine receptor family. 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.

Product Categories/Family for IL12A&IL27B lysate

IL12 signaling pathway

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.