This Human SFRP4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of SFRP4 protein (Cat: 10717-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human sFRP4 (NP_003005.2) (Met 1-Val 346) was expressed with a C-terminal polyhistidine tag.
The recombinant human sFRP4 consists of 336 amino acids after removal of the signal peptide and has a predicted molecular mass of 39 kDa. In SDS-PAGE under reducing conditions, it migrates with an apparent molecular mass of 55-60 kDa due to glycosylation.
Human SFRP4 HEK293 Overexpression Lysate: 用法
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).
安定性 & 保存条件
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.
Human SFRP4 HEK293 Overexpression Lysate: 別名
Human FRP-4 Overexpression Lysate; Human FRPHE Overexpression Lysate; Human sFRP-4 Overexpression Lysate
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. SFRP4 is a member of the SFRP family that contains a cysteine-rich domain homologous to the putative Wnt-binding site of Frizzled proteins called FZ domain and a NTR domain. Mouse SFRP4 is highly expressed in the ovary, and is localized to granulosa cells of periovulatory follicles and corpora lutea. It plays a critical role in placental development and implantation, and is also an important factor in the development of the decidual fibrinoid zone, and in trophoblast apoptosis.
secreted frizzled-related protein 4
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Berndt T., et al., (2003), Secreted frizzled-related protein 4 is a potent tumor-derived phosphaturic agent.J. Clin. Invest. 112:785-794.
Ota T., et al.,(2004), Complete sequencing and characterization of 21,243 full-length human cDNAs.Nat. Genet. 36:40-45.