Category
Neural Stem Cell Growth Factors
Product Name
Human HB-EGF Baculovirus-Insect cells Overexpression Lysate
Synonyms
Human DTR Overexpression Lysate; Human DTS Overexpression Lysate; Human DTSF Overexpression Lysate; Human HEGFL Overexpression Lysate
Molecule Mass
The expressed pro form of human HBEGF consists of 148 amino acids and predicts a molecular mass of 16.4 kDa. The processed soluble HBEGF is a 9.7 kDa protein comprising 86 amino acids generated by proteolytic processing.
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.
Product Description
This Human HB-EGF overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of HB-EGF protein (Cat: 10325-HNAB) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Sequence Information
A DNA sequence encoding the human HBEGF (Q99075) (Met 1-Leu 148) without the C-terminal propeptide (aa 149-208), was expressed and purified.
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).
Application
Western Blot (WB)Optimal dilutions/concentrations should be determined by the end user.