Human PEDF HEK293 Overexpression Lysate Product Information
This Human PEDF overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PEDF protein (Cat: 11104-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human SERPINF1 (NP_002606.3) (Met 1-Pro 418) was fused with a polyhistidine tag at the C-terminus.
The secreted recombinant human SERPINF1 consists of 410 amino acids with the predicted molecular mass of 45.8 kDa as estimated in SDS-PAGE under reducing conditions.
Human PEDF HEK293 Overexpression Lysate Usage Guide
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.
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.
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.
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.
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Human PEDF HEK293 Overexpression Lysate Alternative Names
Human EPC-1 Overexpression Lysate;Human OI12 Overexpression Lysate;Human OI6 Overexpression Lysate;Human PEDF Overexpression Lysate;Human PIG35 Overexpression Lysate
PEDF Background Information
Pigment epithelium-derived factor, also known as PEDF, Serpin F1, and SERPINF1, is a multiple functional protein which has both anti-angiogenic activity and neurotrophic activity at the same time. PEDF is a secreted glycoprotein that belongs to the noninhibitory serpin. It has an alpha/beta core serine-protease inhibitor domain, three major beta-sheets, and ten alpha-helices. PEDF does not inhibit either serine or cysteine proteinases. PEDF exerts diverse physiological activities including anti-angiogenesis, anti-vasopermeability, anti-tumor, and neurotrophic activities. PEDF acts via multiple high affinity ligands and cell receptors. It has been described as a natural angiogenesis inhibitor with neurotrophic and immune-modulation properties. PEDF induces macrophages apoptosis and necrosis through the activation of peroxisome proliferator-activated receptor-gamma by which PEDF could modulate inflammatory reactions in septic shock. It balances angiogenesis in the eye and blocks tumor progression.
serpin peptidase inhibitor, clade F，member 1
Ren, JG. et al., 2005, Med Hypotheses. 64 (1): 74-8. Filleur, S. et al., 2009. J Cell Biochem. 106 (5): 769-75. Kawaguchi, T. et al., 2010, Curr Mol Med. 10 (3): 302-11. Yamagishi, SI. et al., 2010, Curr Mol Med. 10 (3): 284-91. Nakamura, T. et al., 2010, Curr Mol Med. 10 (3): 312-6.