This Human LIFR overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of LIFR protein (Cat: 10628-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of human LIFR (NP_001121143.1) (Met 1-Ser 833) with a C-terminal polyhistidine tag was expressed.
The secreted recombinant human LIFR comprises 800 amino acids with a predicted molecular mass of 91 kDa. As a result of glycosylation, rhLIFR migrates as an approximately 125-135 kDa band in SDS-PAGE under reducing conditions.
Human LIFR HEK293 Overexpression Lysate: 用法
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).
安定性 & 保存条件
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 LIFR HEK293 Overexpression Lysate: 別名
Human CD118 Overexpression Lysate; Human LIF-R Overexpression Lysate; Human SJS2 Overexpression Lysate; Human STWS Overexpression Lysate; Human SWS Overexpression Lysate
LIFR (leukemia inhibitory factor receptor) belongs to the family of cytokine receptors. LIFR forms a high-affinity receptor complex with gp13, which mediates the activity of LIF (leukemia inhibitory factor) and thus affects the differentiation, proliferation, and survival of a wide variety of cells in the adult and the embryo. Besides LIF, LIFR can also bind to and activate CNTF (ciliary neurotrophic factor) and CLC (cardiotrophin like cytokine). Evidence showed that in the retina, LIFR activating LIF, CT-1 and cardiotrophin like cytokine (CLC) are strongly upregulated in response to preconditioning with bright cyclic light leading to robust activation of signal transducer and activator of transcription-3 (STAT3) in a time-dependent manner. Further, blocking LIFR activation during preconditioning using a LIFR antagonist (LIF5) attenuated the induced STAT3 activation and also resulted in reduced preconditioning-induced protection of the retinal photoreceptors. These data demonstrate that LIFR and its ligands play an essential role in endogenous neuroprotective mechanisms triggered by preconditioning-induced stress. LIFR was newly found to be a suppressor of hepatocellular carcinoma (HCC), one of the world's top five causes of cancer-related deaths.
leukemia inhibitory factor receptor alpha
Gearing, D.P. et al.,1991, EMBO J. 10 (10): 2839-2848.
Gearing, D.P. et al.,1992, New Biol. 4 (1): 61-65.
Mosley, B. et al.,1996, J. Biol. Chem. 271 (51): 32635-32643.
Timmermann, A. et al.,2002, Eur. J. Biochem. 269 (11): 2716-2726.
Lass, A. et al.,2002, Fertil. Steril. 76 (6): 1091-1096.