Human PIGR HEK293 Overexpression Lysate

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Human PIGR HEK293 Overexpression Lysate: Product Information

Product Description
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.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
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.
Molecule Mass
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: Usage Guide

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 PIGR HEK293 Overexpression Lysate: Alternative Names

Human FLJ22667 Overexpression Lysate; Human MGC125361 Overexpression Lysate; Human MGC125362 Overexpression Lysate; Human PIGR Overexpression Lysate

PIGR Background Information

Polymeric immunoglobulin receptor, also known asPIGR, is a member of the immunoglobulin superfamilyand aFc 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 ofIgAandIgM. During this process, a cleavage occurs that separates the extracellular (known as the secretory component) from the transmembrane segment of PIGR.
Full Name
polymeric immunoglobulin receptor
References
  • Coyne, R. S. et al., 1995, J. Biol. Chem. 269 (50) :31620-31625. 
  • Kaetzel,C.S., 2001,  Curr Biol. 11(1): R35-38.
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