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The expression of recombinant human COL4A1 protein includes the construction of the expression vector containing the recombinant DNA and the transformation of the expression vector into the E.coli, which provides a variety of macromolecules and components required for transcription and translation. The recombinant DNA was formed by fusing the N-terminal GST tag sequence to the designated sequence encoding the 30-167aa of the human COL4A1 protein. This N-terminal 10xHis-SUMO-tagged and C-terminal Myc-tagged recombinant human COL4A1 protein is also characterized by high purity, >90%. Under SDS-PAGE condition, this recombinant COL4A1 protein migrated to the band of about 40 kDa molecular weight.
Lately, COL4A1 as a novel oncogene associated with the clinical characteristics of malignancy predicts poor prognosis in glioma. As a basement membrane protein, COL4A1 is commonly expressed in many tissues and cell types. The functions of includes extracellular matrix structural constituent; extracellular matrix structural constituent conferring tensile strength; platelet-derived growth factor binding; protein binding. In recent years, variants of COL4A1 have been observed in a spectrum of brain malformations, such as autosomal dominant porencephaly, schizencephaly, intracranial calcifications, small vessel disease causing strokes and cerebral hemorrhages, and hereditary angiopathy with nephropathy, aneurysms, and muscle cramps (HANAC syndrome). The COL4A1-VEGFD fusion leads to production of mature VEGFD after proteolytic processing, which may act as an autocrine growth factor for tumour cells. Besides, many researches pointed that COL4A1 might be associated various diseases conditions like angiopathy, hereditary, nephropathy, aneurysms, and muscle cramps; leukoencephalopathy.
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