Category
Neural Stem Cell Growth Factors
Product Name
Mouse Thy1/CD90 HEK293 Overexpression Lysate
Synonyms
Mouse CD90 Overexpression Lysate; Mouse T25 Overexpression Lysate; Mouse Thy-1 Overexpression Lysate; Mouse Thy-1.2 Overexpression Lysate; Mouse Thy1.1 Overexpression Lysate; Mouse Thy1.2 Overexpression Lysate
Molecule Mass
The recombinant mouse THY1 consists of 123 amino acids and has a predicted molecular mass of 14.2 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rm THY1 is approximately 20-27 kDa due to glycosylation.
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 Mouse Thy1/CD90 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Thy1/CD90 protein (Cat: 50461-M08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the extracellular domain of mouse THY1 (NP_033408.1) without the propeptide (Met 1-Cys 131) was expressed, with a polyhistidine tag at the C-terminus.
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.