Human Frizzled 10 HEK293 Overexpression Lysate 产品信息
Product Description
This Human Frizzled 10 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Frizzled 10 protein (Cat: 16045-H02H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human FZD10 (NP_009128.1) (Met1-Gly161) was expressed with the Fc region of human IgG1 at the C-terminus.
Molecule Mass
The recombinant human FZD10/Fc comprises 379 amino acids and has a predicted molecular mass of 42.8 kDa. The apparent molecular mass of the protein is approximately 48.7 kDa in SDS-PAGE under reducing conditions.
Human Frizzled 10 HEK293 Overexpression Lysate Usage Guide
Preparation Method
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.
Lysis Buffer
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.
Recommend Usage
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.
Sample Buffer
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Human Frizzled 10 HEK293 Overexpression Lysate Alternative Names
Human CD350 Overexpression Lysate;Human FZ-10 Overexpression Lysate;Human Fz10 Overexpression Lysate;Human FzE7 Overexpression Lysate;Human hFz10 Overexpression Lysate
Frizzled 10 Background Information
Frizzled-1, also known as Fz-1, CD35 and FZD1, is a multi-pass membrane protein which belongs to the G-protein coupled receptor Fz/Smo family. Frizzled-1 / FZD1 is abundantly expressed in the cerebellum, followed by cerebral cortex, medulla and spinal cord; very low levels in total brain, frontal lobe, temporal lobe and putamen. It is weakly expressed in adult brain, heart, lung, skeletal muscle, pancreas, spleen and prostate. Frizzled-1 / FZD1 is a receptor for Wnt proteins. Most of frizzled receptors are coupled to the beta-catenin canonical signaling pathway, which leads to the activation of disheveled proteins, inhibition of GSK-3 kinase, nuclear accumulation of beta-catenin and activation of Wnt target genes. A second signaling pathway involving PKC and calcium fluxes has been seen for some family members, it is not yet clear if it represents a distinct pathway or if it can be integrated in the canonical pathway, as PKC seems to be required for Wnt-mediated inactivation of GSK-3 kinase. Both pathways seem to involve interactions with G-proteins. Frizzled-1 / FZD1 may also be involved in transduction and intercellular transmission of polarity information during tissue morphogenesis and/or in differentiated tissues.
Full Name
frizzled class receptor 10
References
Kwon H.S., et al.,(2009), Myocilin is a modulator of Wnt signaling. Mol. Cell. Biol. 29:2139-2154. The MGC Project Team, et al., (2004), The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC).Genome Res. 14:2121-2127. Tanaka S., et al.,(1998), A novel frizzled gene identified in human esophageal carcinoma mediates APC/beta-catenin signals.Proc. Natl. Acad. Sci. U.S.A. 95:10164-10169.
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