|Datasheet||Specific References||Reviews||Related Products||Protocols|
|Baculovirus-Insect Cell lysate that Human MTDH / lyric / metadherin transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).|
|A DNA sequence encoding the human MTDH (AAH45642.1) (Met 1-Thr 582) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.|
|The recombinant human MTDH/GST chimera consists of 819 amino acids and has a calculated molecular mass of 91.7 kDa. It migrates as an approximately 110 kDa band in SDS-PAGE under reducing conditions.|
|Cell lysate was prepared by homogenization 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.|
|Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.|
|12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.|
|Samples are stable for up to twelve months from date of receipt.|
|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.
3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
|1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).|
|Store at 4℃. After re-dissolution, aliquot and store at -80℃.|
|Western blot (WB): Use at an assay dependent dilution.|
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
Metadherin (MTDH), also known as protein LYRIC or astrocyte elevated gene-1 protein (AEG-1), has emerged as an important oncogene protein that is overexpressed in all cancers analyzed so far. MTDH has been demonstrated to play a potential role in several significant aspects of tumor progression. The presence of the lung-homing domain of MTDH on tumor cells at the cell surface where it would be available to bind to vascular targets during metastasis has been confirmed. It has been reported that overexpression of MTDH is associated with progression of disease and poorer prognosis in breast cancer. It has been reported that MTDH overexpression could significantly enhance the invasion and migration of breast cancer cells by inducing epithelial-mesenchymal transition (EMT) that is a process in which polarized epithelial cells are converted into motile mesenchymal cells. Thus, during cancer development, MTDH is conducive to tumor dissemination and metastatic spread.