Human FRZB HEK293 Overexpression Lysate

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Human FRZB HEK293 Overexpression Lysate: Product Information

Product Description
This Human FRZB overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of FRZB protein (Cat: 10370-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human FRZB (NP_001454.2) (Ala32-Asn325) was expressed with a C-terminal polyhistidine tag.
Molecule Mass
The recombinant human FRZB comprises 304 amino acids and has a predicted molecular mass of 34.5 kDa. The apparent molecular mass of the protein is approximately 41 kDa in SDS-PAGE under reducing conditions.

Human FRZB 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 FRZB HEK293 Overexpression Lysate: Alternative Names

Human FRE Overexpression Lysate; Human FRITZ Overexpression Lysate; Human FRP-3 Overexpression Lysate; Human FRZB-1 Overexpression Lysate; Human FRZB-PEN Overexpression Lysate; Human FRZB1 Overexpression Lysate; Human FZRB Overexpression Lysate; Human hFIZ Overexpression Lysate; Human OS1 Overexpression Lysate; Human SFRP3 Overexpression Lysate; Human SRFP3 Overexpression Lysate

FRZB Background Information

FRZB also known as sFRP-3, is a secreted protein containing a domain similar to the putative Wnt-binding region of the frizzled family of transmembrane receptors. FRZB is widely expressed in adult mammalian tissues. In the Xenopus gastrula, FRZB is regulated as a typical Spemann organizer component. FRZB also functions as a competitor for the cell-surface G-protein receptor Frizzled. It is espically important in embryonic development. Defects in FRZB gene can cause female-specific osteoarthritis (OA) susceptibility. FRZB may serve an important role in determining hip shape and may modify the relationship between hip shape and OA.
Full Name
frizzled-related protein
References
  • Hoang B., et al.,(1996), Primary structure and tissue distribution of FRZB, a novel protein related to Drosophila frizzled, suggest a role in skeletal morphogenesis. J. Biol. Chem. 271:26131-26137.
  • Mayr T., et al., (1997), Fritz: a secreted frizzled-related protein that inhibits Wnt activity.Mech. Dev. 63:109-125.
  • Leyns L., et al.,(1997), Frzb-1 is a secreted antagonist of Wnt signaling expressed in the Spemann organizer.Cell 88:747-756.
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