Product Description
This Human Frizzled 4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Frizzled 4 protein (Cat: 16137-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human FZD4 (NP_036325.2) (Met1-Glu180) was expressed with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human FZD4 consists 155 amino acids and predicts a molecular mass of 17.7 kDa.
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 CD344 Overexpression Lysate;Human EVR1 Overexpression Lysate;Human FEVR Overexpression Lysate;Human Fz-4 Overexpression Lysate;Human Fz4 Overexpression Lysate;Human FZD4S Overexpression Lysate;Human FzE4 Overexpression Lysate;Human GPCR Overexpression Lysate;Human hFz4 Overexpression Lysate
Familial exudative vitreoretinopathy (FEVR) is a hereditary blinding disorder that features defects in retinal vascular development. Mutations in FZD4 are known to cause autosomal dominant exudative vitreoretinopathy (EVR1). The mutations in FZD4 are associated with the phenotypes of retinal folds or ectopic macula in FEVR