Human Angiopoietin-4 HEK293 Overexpression Lysate

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Human Angiopoietin-4 HEK293 Overexpression Lysate: Product Information

Product Description
This Human Angiopoietin-4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Angiopoietin-4 protein (Cat: 10293-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human ANGPT4 (Q9Y264) C-terminal fragment (Met 282-Ile 503) was fused with the Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human ANGPT4/Fc chimera is a disulfide-linked homodimeric protein. The reduced monomer consists of 483 amino acids and has a calculated molecular mass of 53.7 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhANGPT4/Fc monomer is approximately 66 kDa due to the glycosylation.

Human Angiopoietin-4 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 Angiopoietin-4 HEK293 Overexpression Lysate: Alternative Names

Human AGP4 Overexpression Lysate; Human ANG-3 Overexpression Lysate; Human ANG4 Overexpression Lysate

Angiopoietin-4 Background Information

Full Name
angiopoietin 4
References
  • Grosios K, et al. (1999) Assignment of ANGPT4, ANGPT1, and ANGPT2 encoding angiopoietins 4, 1 and 2 to human chromosome bands 20p13, 8q22.3?q23 and 8p23.1, respectively, by in situ hybridization and radiation hybrid mapping. Cytogenet. Cell Genet. 84 (1-2): 118-20.
  • Yamakawa M, et al. (2004) Expression of angiopoietins in renal epithelial and clear cell carcinoma cells: regulation by hypoxia and participation in angiogenesis. Am J Physiol Renal Physiol. 287 (4): 649-57.
  • Oliner J, et al. (2004) Suppression of angiogenesis and tumor growth by selective inhibition of angiopoietin-2. Cancer Cell. 6 (5): 507-16.
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