Category
Neural Stem Cell Differentiation Markers
Product Name
Human PDGF alpha receptor/PDGFRA HEK293 Overexpression Lysate
Synonyms
Human CD140A Overexpression Lysate; Human PDGFR-2 Overexpression Lysate; Human PDGFR2 Overexpression Lysate; Human RHEPDGFRA Overexpression Lysate
Molecule Mass
The recombinant human PDGFRα consists of 507 amino acids and predicts a molecular mass of 56.8 kDa. By SDS-PAGE, the apparent molecular mass of rhPDGFRα is approximately 90-100 kDa due to the 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 Human PDGF alpha receptor/PDGFRA overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PDGF alpha receptor/PDGFRA protein (Cat: 10556-HCCH) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
The mature form of human PDGFRa (NP_006197.1) extracellular domain (Met 1-Glu 524) with five amino acids (DDDDK) at the C-terminus 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.