This Human LIF overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of LIF protein (Cat: 14890-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human LIF (P15018) (Met1-Phe202) with a C-terminal polyhistidine tag was expressed.
The recombinant human LIF comprises 191 amino acids and has a predicted molecular mass of 21.2 kDa. The apparent molecular mass of the protein is approximately 35-42 kDa in SDS-PAGE under reducing conditions.
Human LIF HEK293 Overexpression Lysate: 使用指南
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).
稳定性 & 储存条件
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 LIF HEK293 Overexpression Lysate: 别称
Human CDF Overexpression Lysate; Human DIA Overexpression Lysate; Human HILDA Overexpression Lysate; Human Leukemia Inhibitory Factor Overexpression Lysate; Human MLPLI Overexpression Lysate
Leukemia inhibitory factor (LIF) is a pleiotropic glycoprotein belonging to the IL-6 family of cytokines. It is involved in growth promotion and cell differentiation of different types of target cells, influence bone metabolism, cachexia, neural development, embryogenesis, and inflammation. LIF has potent proinflammatory properties, being the inducer of the acute phase protein synthesis and affecting cell recruitment into the area of damage or inflammation. LIF is also one of the cytokines that are capable to regulate the differentiation of embryonic stem cells, hematopoietic, and neuronal cells. LIF binds to the specific LIF receptor (LIFR-α) which forms a heterodimer with a specific subunit common to all members of that family of receptors, the GP130 signal-transducing subunit. This leads to the activation of the JAK/STAT and MAPK cascades. Due to its polyfunctional activities, LIF is involved in the pathogenic events and development of many diseases of various origins.
leukemia inhibitory factor
Salas EM, et al. (2011) LIF, a Novel STAT5-Regulated Gene, Is Aberrantly Expressed in Myeloproliferative Neoplasms. Genes Cancer. 2 (5): 593-6.
Chodorowska G, et al. (2004) Leukemia inhibitory factor (LIF) and its biological activity. Ann Univ Mariae Curie Sklodowska Med. 59 (2): 189-93.
Garcia-Campana AM, et al. (2007) LIF detection of peptides and proteins in CE. Electrophoresis. 28 (1-2): 208-32.