Human TCCR HEK293 Overexpression Lysate: Product Information
This Human TCCR overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TCCR protein (Cat: 10489-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Arg 514) of human IL27Rα (NP_004834.1) was fused with the Fc region of human IgG1 at the C-terminus.
The recombinant human IL27Rα/Fc is a disulfide-linked homodimeric protein after removal of the signal peptide. The reduced monomer consists of 722 amino acids and has a predicted molecular mass of 79 kDa. As a result of glycosylation, rh IL27Rα/Fc monomer migrates as an approximately 105-115 kDa band in SDS-PAGE under reducing conditions.
Human TCCR 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 TCCR HEK293 Overexpression Lysate: Alternative Names
Human CRL1 Overexpression Lysate; Human IL-27RA Overexpression Lysate; Human IL27R Overexpression Lysate; Human TCCR Overexpression Lysate; Human WSX1 Overexpression Lysate; Human zcytor1 Overexpression Lysate
TCCR Background Information
The interleukin-27 receptor is a type I cytokine receptor for interleukin-27. It is a heterodimer composed of the interleukin 27 receptor, alpha subunit and glycoprotein 13. WSX-1/IL-27R, a class I cytokine receptor that is homologous to the β2 chain of the IL-12R in both sequence and structure, is highly expressed by resting/naive CD4+ T cells and CD8+ T cells. WSX-1/IL-27R belongs to the IL-6/IL-12 family of cytokines and induced proliferation of naive CD4(+) T cells and the generation of a Th1-type adaptive immune response. WSX-1/IL-27R functions as a receptor for IL27. IL-27 not only contributes to the development of an adaptive immune response through its action on CD4(+) T cells, it also directly acts on cells of the innate immune system. WSX-1/IL-27R requires IL6ST/gp13 to mediate signal transduction in response to IL27. This signaling system acts through STAT3 and STAT1. It is involved in the regulation of Th1-type immune responses. IL-27R also appears to be involved in innate defense mechanisms. IL27RA is essential for transcriptional activation of STAT1 and for augmenting the induction of T-bet expression during initiation of Th1 cell differentiation.
interleukin 27 receptor, alpha
Pflanz S, et al.. (2004) WSX-1 and glycoprotein 130 constitute a signal-transducing receptor for IL-27. J Immunol. 172(4): 2225-31.
Yoshida H, et al.. (2001) WSX-1 is required for the initiation of Th1 responses and resistance to L. major infection. Immunity. 215(4): 569-78.
Villarino A, et al.. (2003) The IL-27R (WSX-1) is required to suppress T cell hyperactivity during infection. Immunity. 19(5): 645-55.
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