Tie-1 Overexpression Lysate

Western Blot: TIE1 Overexpression Lysate (Adult Normal) [NBL1-16909] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for TIE1.

• Reactivity: Hu
• Applications: WB

Tie-1 Overexpression Lysate Summary

• Description:

  Tie-1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC207771
  
  Accession#: NM_005424
  
  Protein Tag: C-MYC/DDK
  
  You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: TIE1

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 124.9 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for Tie-1 Overexpression Lysate

• EC 2.7.10
• EC 2.7.10.1
• JTK14soluble TIE1 variant 3
• receptor tyrosine kinase
• soluble TIE1 variant 1
• soluble TIE1 variant 2
• soluble TIE1 variant 5
• Tie1
• Tie-1
• TIEsoluble TIE1 variant 4
• tyrosine kinase with immunoglobulin and epidermal growth factor homologydomains 1
• tyrosine kinase with immunoglobulin-like and EGF-like domains 1
• tyrosine-protein kinase receptor Tie-1

Background

TIE1/TIE (tyrosine kinase with Ig and EGF homology domains 1) and TIE2/Tek define a new class of the receptor tyrosine kinase (RTK) subfamily with unique structural characteristics: two immunoglobulin like domains flanking three epidermal growth factor (EFG) like domains followed by three fibronectin type III like repeats in the extracellular region and a split tyrosine kinase domain in the cytoplasmic region. Human TIE1 cDNA encodes a 1138 amino acid residue precursor protein with a putative signal peptide, an extracellular domain, and a cytoplasmic domain. Human TIE1/Fc, a disulfide linked homodimeric protein, has a calculated molecular mass of approximately 107 kDa. Due to glycosylation, the protein migrates to approximately 160 kDa in SDS PAGE under reducing conditions. TIE1 and TIE2, expressed primarily on endothelial and hematopoietic progenitor cells, play important roles in angiogenesis, vasculogenesis, and hematopoiesis. In developing vascular endothelial cells, TIE1 and TIE2 are specifically expressed. Two ligands that bind TIE have been identified, angiopoietin 1 and angiopoietin 2. Based on gene targeting studies, the in vivo functions of TIE1 are related to endothelial cell differentiation. The receptor tyrosine kinase TIE also plays a role in the survival and integrity of the endothelium.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: TIE1