This Human GDF15 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of GDF15 protein (Cat: 10936-H01H) from the overexpression lysate was verified.
A DNA sequence encoding the mature form of human GDF15 (P01857.1) (Ala 197-Ile 308) was expressed with the fused Fc region of human IgG1 at the N-terminus.
The recombinant human Fc/GDF15 is a disulfide-linked homodimeric protein. The reduced monomer consists of 370 amino acids and has a predicted molecular mass of 40.6 kDa. rhGDF15/Fc monomer migrates as an approximately 44 kDa protein in SDS-PAGE under reducing conditions.
Human GDF15 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 GDF15 HEK293 Overexpression Lysate: 别称
Human GDF-15 Overexpression Lysate; Human MIC-1 Overexpression Lysate; Human MIC1 Overexpression Lysate; Human NAG-1 Overexpression Lysate; Human PDF Overexpression Lysate; Human PLAB Overexpression Lysate; Human PTGFB Overexpression Lysate
Growth-differentiation factor 15 (GDF15), also known as MIC-1, is a secreted member of the transforming growth factor (TGF)-β superfamily, as a novel antihypertrophic regulatory factor in the heart. GDF-15 / GDF15 is not expressed in the normal adult heart but is induced in response to conditions that promote hypertrophy and dilated cardiomyopathy and it is expressed highly in liver. GDF-15 / GDF15 has a role in regulating inflammatory and apoptotic pathways in injured tissues and during disease processes. GDF-15 / GDF15 is synthesized as precursor molecules that are processed at a dibasic cleavage site to release C-terminal domains containing a characteristic motif of 7 conserved cysteines in the mature protein. GDF-15 / GDF15 overexpression arising from an expanded erythroid compartment contributes to iron overload in thalassemia syndromes by inhibiting hepcidin expression.
growth differentiation factor 15
Ago T, et al. (2006) GDF15, a cardioprotective TGF-beta superfamily protein. Circ Res. 98 (3): 294-297.
Hsiao E, et al. (2000) Characterization of growth-differentiation factor 15, a transforming growth factor beta superfamily member induced following liver injury. Mol Cell Biol. 20 (10): 3742-51.
Zimmers T, et al. (2005) Growth differentiation factor-15/macrophage inhibitory cytokine-1 induction after kidney and lung injury. Shock. 23 (6): 543-8.