Human Periostin HEK293 Overexpression Lysate 产品信息
Product Description
This Human Periostin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Periostin protein (Cat: 10299-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human POSTN isoform 3 (NP_001129407.1) corresponding to amino acid (Met 1-Gln 781) was expressed with a C-terminal polyhistidine tag.
Molecule Mass
The recombinant human POSTN consists of 771 amino acids and has a predicted molecular mass of 86 kDa. In SDS-PAGE under reducing conditions, it migrates with an apparent molecular mass of 80 kDa due to glycosylation.
Human Periostin 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 Periostin HEK293 Overexpression Lysate Alternative Names
Human OSF-2 Overexpression Lysate;Human OSF2 Overexpression Lysate;Human PDLPOSTN Overexpression Lysate;Human periostin Overexpression Lysate;Human PN Overexpression Lysate
Periostin Background Information
Periostin ( POSTN ), also known as OSF2 (osteoblast specific factor 2), is a heterofunctional secreted extracellular matrix (ECM) protein comprised of four fasciclin domains that promotes cellular adhesion and movement, as well as collagen fibrillogenesis. Postn is expressed in unique growth centers during embryonic development where it facilitates epithelial-mesenchymal transition (EMT) of select cell populations undergoing reorganization. In the adult, Postn expression is specifically induced in areas of tissue injury or areas with ongoing cellular re-organization. In the adult heart Postn is induced in the ventricles following myocardial infarction, pressure overload stimulation, or generalized cardiomyopathy. Although the detailed function of Postn is still unclear, Postn-integrin interaction is thought to be involved in tumor development. Postn is frequently overexpressed in various types of human cancers, stimulating metastatic growth by promoting cancer cell survival, invasion and angiogenesis, and can be a useful marker to predict the behavior of cancer.
Full Name
periostin, osteoblast specific factor
References
Kudo,Y. et al., 2007, Histol Histopathol. 22 (10):1167-1174. Li, J.S. et al., 2007, World J Gastroenterol. 13 (39): 5261-5266. Oku, E. et al., 2008, Int J Hematol. 88 (1): 57-63. Hamilton, D.W. et al., 2008, J Cell Commun Signal. 2(1-2):9-17. Puglisi, F.J et al., 2008, Clin Pathol. 61 (4): 494-498. Conway, S. J. et al., 2008, Curr Genomics. 9 (8): 548-555.