This Mouse TROP2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TROP2 protein (Cat: 50922-M02H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of mouse TACSTD2 (Q8BGV3?) (Met 1-Gln 270) was fused with the Fc region of human IgG1 at the C-terminus.
The secreted recombinant mouse TACSTD2/Fc is a disulfide-linked homodimer. The reduced monomer comprises 487 amino acids and has a calculated molecular mass of 55 kDa. As a result of glycosylation, the apparent molecular mass of rmTACSTD2/Fc monomer is approximately 66 kDa in SDS-PAGE under reducing conditions.
Mouse TROP2 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.
TROP-2, also referred to as tumor-associated calcium signal transducer 2 (TACSTD2), GA733-1 or M1S1, is a cell surface glycoprotein highly expressed in a wide variety of epithelial cancers. In contrast, there is little or no expression of Trop-2 in adult somatic tissue. Because it is a cell surface protein that is selectively expressed in tumor cells, Trop-2 is a potential therapeutic target. The cytoplasmic tail of Trop-2 possesses potential serine and tyrosine phosphorylation sites and a phosphatidyl-inositol binding consensus sequence. Trop-2 transduces an intracellular calcium signal, which are consistent with the hypothesis that it acts as a cell surface receptor and support a search for a physiological ligand. TROP2 encoding by an intronless gene was originally defined by the monoclonal antibody GA733, and is a member of a family of at least two type I membrane proteins. The other known member is GA733-2, also called EpCAM and TROP1. It has been suggested by studies that the GA733-1 gene was formed by the retroposition of the GA733-2 gene via an mRNA intermediate.