Human CDO HEK293 Overexpression Lysate

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Human CDO HEK293 Overexpression Lysate: Product Information

Product Description
This Human CDO overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CDO protein (Cat: 11965-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human CDON (AAI14436.1) extracellular domain (Met 1-Asp 963) was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human CDON consists of 949 amino acids and has a calculated molecular mass of 103 kDa. It migrates as an approximately 125 kDa band in SDS-PAGE under reducing conditions.

Human CDO 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 CDO HEK293 Overexpression Lysate: Alternative Names

Human CDO Overexpression Lysate; Human CDON1 Overexpression Lysate; Human HPE11 Overexpression Lysate; Human ORCAM Overexpression Lysate

CDO Background Information

Cell adhesion molecule-related, down-regulated by oncogenes (CDON), also known as CDO, is an Ig superfamily member, is a component of a cell surface receptor that positively regulates skeletal myogenesis. Brother of CDO (BOC) is a cell surface receptor that derives its name from the structurally related protein, CDON. They are components of a cell surface receptor that positively regulates myogenesis in vitro. Expression of Cdo and Boc in myoblast cell lines is downregulated by the ras oncogene, and forced re-expression of either Cdo or Boc can override ras-induced inhibition of myogenic differentiation. CDO and BOC play a role in the inverse relationship between differentiation and transformation of cells in the skeletal muscle lineage. CDON binds to Bnip-2 and JLP, scaffold proteins for Cdc42 and p38alpha/beta MAPK, respectively. The Bnip-2/Cdc42 and JLP/p38alpha/beta complexes associate in a CDON-dependent manner, resulting in Bnip-2/Cdc42-dependent p38alpha/beta activation and stimulation of cell differentiation. It is proposed that CDO mediates, at least in part, the effects of cell-cell interactions between muscle precursors that are critical in myogenesis.
Full Name
cell adhesion associated, oncogene regulated
References
  • Kang JS, et al. (1998) CDO, a robo-related cell surface protein that mediates myogenic differentiation. J Cell Biol. 143(2): 403-13.
  • Wegorzewska M, et al. (2003) Overexpression of the immunoglobulin superfamily members CDO and BOC enhances differentiation of the human rhabdomyosarcoma cell line RD. Mol Carcinog. 37(1): 1-4.
  • Cole F, et al. (2003) Microform holoprosencephaly in mice that lack the Ig superfamily member Cdon. Curr Biol. 13(5): 411-5.
  • Jehee FS, et al. (2006) Mutational screening of FGFR1, CER1, and CDON in a large cohort of trigonocephalic patients. Cleft Palate Craniofac J. 43(2): 148-51.
  • Kavran JM, et al. (2010) All mammalian Hedgehog proteins interact with cell adhesion molecule, down-regulated by oncogenes (CDO) and brother of CDO (BOC) in a conserved manner. J Biol Chem. 285(32): 24584-90.
  • Lu M, et al. (2010) N-cadherin ligation, but not Sonic hedgehog binding, initiates Cdo-dependent p38alpha/beta MAPK signaling in skeletal myoblasts. Proc Natl Acad Sci U S A. 107(9): 4212-7.
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