This Human ErbB4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ErbB4 protein (Cat: 10363-H03H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Pro 651) of human ErbB4 precursor (NP_001036064.1) was fused with the C-terminal His-tagged Fc region of human IgG1 at the C-terminus.
The recombinant human ErbB4/Fc is a disulfide-linked homodimeric protein after proteolytic removal of the signal peptide. The reduced monomer consists of 871 amino acids and predicts a molecular mass of 97.5 kDa. As a result of glycosylation, the rh ErbB4/Fc monomer migrates as approximately 125-135 kDa band in SDS-PAGE under reducing conditions.
Human ErbB4 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 ErbB4 HEK293 Overexpression Lysate: 别称
Human ALS19 Overexpression Lysate; Human HER4 Overexpression Lysate; Human p180erbB4 Overexpression Lysate
ERBB4 is a single-pass type I membrane protein with multiple cysteine rich domains, a transmembrane domain, a tyrosine kinase domain, a phosphotidylinositol-3 kinase binding site and a PDZ domain binding motif. ERBB4 is expressed at highest levels in brain, heart, kidney, in addition to skeletal muscle, parathyroid, cerebellum, pituitary, spleen, testis and breast. And lower levels in thymus, lung, salivary gland, and pancreas. It specifically binds to and is activated by neuregulins, NRG-2, NRG-3, heparin-binding EGF-like growth factor, betacellulin and NTAK. ERBB4 also can be activated by other factors and induces a variety of cellular responses including mitogenesis and differentiation. ERBB4 regulates development of the heart, the central nervous system and the mammary gland, gene transcription, cell proliferation, differentiation, migration and apoptosis. It is required for normal cardiac muscle differentiation during embryonic development, and for postnatal cardiomyocyte proliferation. ERBB4 also play a role on the normal development of the embryonic central nervous system, especially for normal neural crest cell migration and normal axon guidance. It is required for mammary gland differentiation, induction of milk proteins and lactation.