Product Description
This Human PSG1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PSG1 protein (Cat: 15378-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human PSG1 (NP_001171754.1) (Met1-Pro419) was expressed with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human PSG1 consists 396 amino acids and predicts a molecular mass of 44.9 kDa.
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).
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Human B1G1 Overexpression Lysate;Human CD66f Overexpression Lysate;Human DHFRP2 Overexpression Lysate;Human FL-NCA-1/2 Overexpression Lysate;Human PBG1 Overexpression Lysate;Human PS-beta-C/D Overexpression Lysate;Human PS-beta-G-1 Overexpression Lysate;Human PSBG-1 Overexpression Lysate;Human PSBG1 Overexpression Lysate;Human PSG95 Overexpression Lysate;Human PSGGA Overexpression Lysate;Human PSGIIA Overexpression Lysate;Human SP1 Overexpression Lysate
Pregnancy-specific glycoproteins (PSGs) are a family of Ig-like proteins secreted by specialized placental cells. The PSG1 structure is composed of a single Ig variable region-like N-terminal domain and three Ig constant region-like domains termed A1, A2, and B2. Members of the human and murine PSG family have been shown to induce anti-inflammatory cytokines from monocytes and macrophages and to stimulate angiogenesis.Two distinct domains of PSG1 are involved in the regulation of TGF-beta1 and provide a mechanistic framework for how PSGs modulate the immunoregulatory environment at the maternal-fetal interface for successful pregnancy outcome.