Mouse SFRP1 CHO Stable Overexpression Lysate: 产品信息
This Mouse SFRP1 overexpression lysate was created in CHO Stable Cells and intented for use as a Western blot (WB) positive control. Purification of SFRP1 protein (Cat: 50033-M08S) from the overexpression lysate was verified.
CHO Stable Cells
A DNA sequence encoding the mouse SFRP1 (AAC53145.1) (Met1-Lys314) was expressed with a polyhistidine tag at the C-terminus.
The recombinant mouse SFRP1 consists of 283 amino acids and predicts a molecular mass of 32.6 kDa.
Mouse SFRP1 CHO Stable Overexpression Lysate: 使用指南
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.
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).
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.
Secreted frizzled-related protein 1, also known as sFRP1, is a 35 kDa prototypical member of the SFRP family. SFRP family consists of five secreted glycoproteins in humans acting as extracellular signaling ligands. Each is approximately 3 amino acids in length and contains a cysteine-rich domain (CRD) that shares 3-5% sequence homology with the CRD of Frizzled (Fz) receptors, a putative signal sequence, and a conserved hydrophilic carboxy-terminal domain. SFRPs act as soluble modulators of Wnt signaling, counteracting Wnt-induced effects at high concentrations and promoting them at lower concentrations. SFRPs are able to bind Wnt proteins and Fz receptors in the extracellular compartment. The interaction between SFRPs and Wnt proteins prevents the latter from binding the Fz receptors. The Wnt pathway plays a key role in embryonic development, cell differentiation and cell proliferation. The deregulation of this critical developmental pathway occurs in several human tumor entities. Mouse sFRP1 is highly expressed in kidney and embryonic heart, as well as in the eye, where it is principally localized to the ciliary body and the lens epithelium.
secreted frizzled-related protein 1
Finch P.W., et al.,(1997), Purification and molecular cloning of a secreted, Frizzled-related antagonist of Wnt action. Proc. Natl. Acad. Sci. U.S.A. 94:6770-6775.
Melkonyan H.S., et al., (1997), SARPs: a family of secreted apoptosis-related proteins.Proc. Natl. Acad. Sci. U.S.A. 94:13636-13641.
Zhou Z., et al.,(1998), Up-regulation of human secreted frizzled homolog in apoptosis and its down-regulation in breast tumors.Int. J. Cancer 78:95-99.