Human OMGP HEK293 Overexpression Lysate

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Human OMGP HEK293 Overexpression Lysate: Product Information

Product Description
This Human OMGP overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of OMGP protein (Cat: 10269-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the mature form of human OMGP (P23515-1) (Met 1-Pro 416) without the pro peptide was expressed, fused with a polyhistidine tag at the C-terminus.
Molecule Mass
The secreted recombinant human OMGP comprises 403 amino acids and has a predicted molecular mass of 46 kDa. The apparent molecular mass of rh OMGP is approximately 120-130 kDa in SDS-PAGE under reducing conditions due to high glycosylation.

Human OMGP 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 OMGP HEK293 Overexpression Lysate: Alternative Names

Human OMGP Overexpression Lysate

OMGP Background Information

Oligodendrocyte-myelin glycoprotein, also known as OMG and OMGP, is a cell membrane protein which contains eightLRR (leucine-rich) repeats. OMG / OMGP is a glycosylphosphatidylinositol-anchored protein expressed by neurons and oligodendrocytes in the central nervous system (CNS). OMG / OMGP is a cell adhesion molecule contributing to the interactive process required for myelination in the central nervous system. OMG / OMGP play roles in both the developing and adult central nervous system. OMG / OMGP participats in growth cone collapse and inhibition of neurite outgrowth through its interaction with NgR, the receptor for Nogo. This function requires its leucine-rich repeat domain, a highly conserved region in OMgp during mammal evolution. OMG / OMGP leucine-rich repeat domain is also implicated in the inhibition of cell proliferation. OMG / OMGP may also be involved in the formation and maintenance of myelin sheaths. Cell proliferation, neuronal sprouting and myelination are crucial processes involved in brain development and regeneration after injury.
Full Name
oligodendrocyte myelin glycoprotein
References
  • Viskochil D., et al.,(1991), The gene encoding the oligodendrocyte-myelin glycoprotein is embedded within the neurofibromatosis type 1 gene. Mol. Cell. Biol. 11:906-912.
  • Mikol D.D., et al., (1990), The oligodendrocyte-myelin glycoprotein belongs to a distinct family of proteins and contains the HNK-1 carbohydrate.J. Cell Biol. 110:471-479.
  • Mikol D.D., et al.,(1990), Structure and chromosomal localization of the gene for the oligodendrocyte-myelin glycoprotein.J. Cell Biol. 111:2673-2679.
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