Human EGFL6 Baculovirus-Insect cells Overexpression Lysate

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Human EGFL6 Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human EGFL6 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of EGFL6 protein (Cat: 10867-H08B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human EGFL6 (NP_056322.2)(Met1-Asp553) was expressed with a C-terminal polyhistidine tag.
Molecule Mass
The secreted recombinant human EGFL6 consists of 542 amino acids and predicts a molecular mass of 60.5 KDa. The apparent molecular mass of the protein is approximately 62 KDa in SDS-PAGE under reducing conditions due to glycosylation.

Human EGFL6 Baculovirus-Insect cells 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 EGFL6 Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human MAEG Overexpression Lysate; Human W80 Overexpression Lysate

EGFL6 Background Information

EGFL6, also known as EGF-L6, belongs to the epidermal growth factor (EGF) repeat superfamily. Members of this superfamily are characterized by the presence of EGF-like repeats and are often involved in the regulation of cell cycle, proliferation, and developmental processes. EGFL6 contains a signal peptide, suggesting that it is secreted; an EGF repeat region consisting of 4 complete EGF-like repeats and 1 partial EGF-like repeat, 3 of which have a calcium-binding consensus sequence; an arg-gly-asp integrin association motif; and a MAM domain, which is believed to have an adhesive function. EGFL6 gene is expressed early during development, and its expression has been detected in lung and meningioma tumors.
Full Name
EGF-like-domain, multiple 6
References
  • Kimura K. et al., 2006, Genome Res. 16 (1): 55-65.
  • Lee SH. et al., 2008, Pathology. 40 (1): 95-7.
  • Oberauer R. et al., 2010, Mol Cell Biochem. 343 (1-2): 257-69.
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