Human TGF beta 1 HEK293 Overexpression Lysate: Product Information
This Human TGF beta 1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TGF beta 1 protein (Cat: 10804-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the full length of human TGFβ1 (NP_000651.3) (Met 1-Ser 390) was fused with a C-terminal polyhistidine tag.
The recombinant human latent TGFβ1 consists of 370 amino acids and has a calculated molecular mass of 42.4 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh TGFβ1 is approximately 16, 38 and 55 kDa corresponding to mature TGFβ1, LAP protein and inacitve latent TGFβ1 respectively due to glycosylation. In non-reduced SDS-PAGE, it migrates as an approximately 110 kDa protein consisting of a TGFβ1 homodimer non-covalently linked to a LAP homodimer.
Human TGF beta 1 HEK293 Overexpression Lysate: Usage Guide
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).
Stability & Storage
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 TGF beta 1 HEK293 Overexpression Lysate: Alternative Names
Human CED Overexpression Lysate; Human DPD1 Overexpression Lysate; Human LAP Overexpression Lysate; Human TGF-beta 1 Overexpression Lysate; Human TGFB Overexpression Lysate; Human TGFbeta Overexpression Lysate
TGF beta 1 Background Information
TGF-beta 1 is a member of the transforming growth factor beta (TGF-beta) family. The transforming growth factor-beta family of polypeptides are involved in the regulation of cellular processes, including cell division, differentiation, motility, adhesion and death. TGF-beta 1 positively and negatively regulates many other growth factors. It inhibits the secretion and activity of many other cytokines including interferon-γ, tumor necrosis factor-alpha and various interleukins. It can also decrease the expression levels of cytokine receptors. Meanwhile, TGF-beta 1 also increases the expression of certain cytokines in T cells and promotes their proliferation, particularly if the cells are immature. TGF-beta 1 also inhibits proliferation and stimulates apoptosis of B cells, and plays a role in controlling the expression of antibody, transferrin and MHC class II proteins on immature and mature B cells. As for myeloid cells, TGF-beta 1can inhibit their proliferation and prevent their production of reactive oxygen and nitrogen intermediates. However, as with other cell types, TGF-beta 1 also has the opposite effect on cells of myeloid origin. TGF-beta 1 is a multifunctional protein that controls proliferation, differentiation and other functions in many cell types. It plays an important role in bone remodeling as it is a potent stimulator of osteoblastic bone formation, causing chemotaxis, proliferation and differentiation in committed osteoblasts. Once cells lose their sensitivity to TGF-beta1-mediated growth inhibition, autocrine TGF-beta signaling can promote tumorigenesis. Elevated levels of TGF-beta1 are often observed in advanced carcinomas, and have been correlated with increased tumor invasiveness and disease progression.
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