Human ALK4/ACVR1B HEK293 Overexpression Lysate: 产品信息
This Human ALK4/ACVR1B overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ALK4/ACVR1B protein (Cat: 10583-H03H) from the overexpression lysate was verified.
A DNA sequence encoding the human ACVR1B (NP_004293.1) extracellular domain (Met 1-Glu 126) was fused with C-terminal His-tagged Fc region of human IgG1 at the C-terminus.
The recombinant human ALK4/Fc is a disulfide-linked homodimer. The reduced monomer consists of 351 amino acids and has a predicted molecular mass of 39.6 kDa. In SDS-PAGE under reducing conditions, rhALK4/Fc monomer migrates as an approximately 46 kDa band due to glycosylation, with ~15% free Fc fragment.
Human ALK4/ACVR1B 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 ALK4/ACVR1B HEK293 Overexpression Lysate: 别称
Human ACTRIB Overexpression Lysate; Human ACVRLK4 Overexpression Lysate; Human ALK4 Overexpression Lysate; Human SKR2 Overexpression Lysate
ALK-4 (Activin Receptor-Like Kinase 4) or ACVR1B (Activin A Receptor, type 1B), belongs to the protein kinase superfamily, TKL Ser/Thr protein kinase family, and TGFB receptor subfamily. ALK-4/ACVR1B acts as a transducer of activin or activin like ligands signals. Activin binds to either ACVR2A or ACVR2B and then forms a complex with ACVR1B. The known type II activin receptors include ActRII and ActRIIB, while the main type I activin receptor in mammalian cells is ALK-4 (ActRIB). In the presence of activin, type II and type I receptors form complexes whereby the type II receptors activate ALK-4 through phosphorylation. The activated ALK-4, in turn, transduces signals downstream by phosphorylation of its effectors, such as Smads, to regulate gene expression and affect cellular phenotype. ALK-4/ACVR1B is an important regulator of vertebrate development, with roles in mesoderm induction, primitive streak formation, gastrulation, dorsoanterior patterning, and left-right axis determination.
activin A receptor, type IB
Chen Y, et al. (2005) Developmental analysis of activin-like kinase receptor-4 (ALK4) expression in Xenopus laevis. 232(2): 393-8.
J. Massagu. (1998) TGF- SIGNAL TRANSDUCTION. Annual Review of Biochemistry. 67: 753-91.