Human gp130/IL6ST HEK293 Overexpression Lysate: Product Information
This Human gp130/IL6ST overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of gp130/IL6ST protein (Cat: 10974-HCCH) from the overexpression lysate was verified.
A DNA sequence encoding the human IL6RB (NP_002175.2)(Met1-Ile618) was expressed with six amino acids (ENLYFQ) at the C-terminus.
The recombinant human IL6RB consists of 603 amino acids and predicts a molecular mass of 68.6 KDa. It migrates as an approximately 69-99 KDa band in SDS-PAGE under reducing conditions due to glycosylation.
Human gp130/IL6ST 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 gp130/IL6ST HEK293 Overexpression Lysate: Alternative Names
Human CD130 Overexpression Lysate; Human CDW130 Overexpression Lysate; Human GP130 Overexpression Lysate; Human IL-6RB Overexpression Lysate
gp130/IL6ST Background Information
Glycoprotein 13 (also known as gp13, IL6ST, IL6-beta or CD13) is a transmembrane protein which is the founding member of the class of all cytokine receptors. CD13/gp13 is a signal transducer shared by many cytokines, including interleukin 6 (IL6), ciliary neurotrophic factor (CNTF), leukemia inhibitory factor (LIF), and Oncostatin M (OSM). CD13/gp13 functions as a part of the cytokine receptor complex. The activation of this protein is dependent upon the binding of cytokines to their receptors. CD13/gp13 plays a critical role in regulating myocyte apoptosis. Alternatively spliced transcript variants encoding distinct isoforms have been described. A related pseudogene has been identified on chromosome 17. The receptor systems for IL6, LIF, OSM, CNTF, IL11, CTF1 and BSF3 can utilize gp13 for initiating signal transmission. CD13/gp13 binds to IL6/IL6R (alpha chain) complex, resulting in the formation of high-affinity IL6 binding sites, and transduces the signal. CD13/gp13 may have a role in embryonic development. The type I OSM receptor is capable of transducing OSM-specific signaling events.
interleukin 6 signal transducer
Hibi, et al. (1990) Molecular cloning and expression of an IL-6 signal transducer, gp130. Cell. 63 (6): 1149-57.
Kim H, et al. (1997) Transmembrane domain of gp130 contributes to intracellular signal transduction in hepatic cells. J Biol Chem. 272 (49): 30741-7.
Giordano V, et al. (1997) Shc mediates IL-6 signaling by interacting with gp130 and Jak2 kinase. J Immunol. 158 (9): 4097-103.
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