This Mouse DOPA Decarboxylase/DDC overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of DOPA Decarboxylase/DDC protein from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Mouse
Sequence Information
A DNA sequence encoding the mouse DDC (O88533) (Met 1-Glu 480) was expressed,with a C-terminal polyhistidine tag.
Molecule Mass
The recombinant mouse DDC consists of 490 amino acids and has a calculated molecular mass of 55.2 kDa. It migrates as an approximately 50 kDa band in SDS-PAGE under reducing conditions.
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.
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.
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).
Synonyms
Mouse Aadc Overexpression Lysate
Application
Western Blot (WB). Optimal dilutions/concentrations should be determined by the end user.
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