Human SFRP1 HEK293 Overexpression Lysate: Product Information
This Human SFRP1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of SFRP1 protein (Cat: 10680-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human sFRP1 (NP_003003.3) (Met 1-Lys 314) was expressed with a C-terminal polyhistidine tag.
The recombinant human sFRP1 consists of 294 amino acids after removal of the signal peptide and has a predicted molecular mass of 34 kDa. In SDS-PAGE under reducing conditions, it migrates with an apparent molecular mass of 38 kDa due to glycosylation.
Human SFRP1 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 SFRP1 HEK293 Overexpression Lysate: Alternative Names
Human FRP Overexpression Lysate; Human FRP-1 Overexpression Lysate; Human FRP1 Overexpression Lysate; Human FrzA Overexpression Lysate; Human SARP2 Overexpression Lysate
SFRP1 Background Information
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 humansacting 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 ofFrizzled(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 bindWntproteins 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.
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