This Human PIGR overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PIGR protein (Cat: 10131-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Arg 638, 365 Ser/Gly) of human PIGR (NP_002635.2) was expressed with a C-terminal polyhistidine tag.
The soluble form of recombinant human PIGR consists of 630 amino acids after removal of the signal peptide and has a predicted molecular mass of 69 kDa. In SDS-PAGE under reducing conditions, it migrates with an apparent molecular mass of 80-90 kDa due to glycosylation.
Human PIGR HEK293 Overexpression Lysate: 使用指南
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.
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).
稳定性 & 储存条件
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 PIGR HEK293 Overexpression Lysate: 别称
Human FLJ22667 Overexpression Lysate; Human MGC125361 Overexpression Lysate; Human MGC125362 Overexpression Lysate; Human PIGR Overexpression Lysate
Polymeric immunoglobulin receptor, also known as PIGR, is a member of the immunoglobulin superfamily and a Fc receptor. The ectodomain of this receptor consists of five units with homology to the variable units of immunoglobulins and a transmembrane region, which also has some homology to certain immunoglobulin variable regions. PIGR is expressed on several glandular epithelia including those of liver and breast. The deduced amino-acid sequence has a length of 764 residues and shows an overall similarity of 56% and 64% with the rabbit and rat counterpart. PIGR mediates transcellular transport of polymeric immunoglobulin molecules, and thus facilitates the secretion of IgA and IgM. During this process, a cleavage occurs that separates the extracellular (known as the secretory component) from the transmembrane segment of PIGR.
polymeric immunoglobulin receptor
Coyne, R. S. et al., 1995, J. Biol. Chem. 269 (50) :31620-31625.