eIF3e Overexpression Lysate

Western Blot: eIF3e Overexpression Lysate (Adult Normal) [NBL1-10192] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for eIF3e.

• Reactivity: Hu
• Applications: WB

eIF3e Overexpression Lysate Summary

• Description:

  eIF3e Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC204788
  
  Accession#: NM_001568
  
  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: EIF3E

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: 52 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 eIF3e Overexpression Lysate

• eIF-3 p48
• eIF3e
• eIF3-p48
• EIF3S6EIF3-P48
• Eukaryotic translation initiation factor 3 subunit 6
• eukaryotic translation initiation factor 3 subunit E
• eukaryotic translation initiation factor 3, subunit 6 (48kD)
• eukaryotic translation initiation factor 3, subunit 6 48kDa
• eukaryotic translation initiation factor 3, subunit E
• INT6eIF3-p46
• mammary tumor-associated protein INT6
• murine mammary tumor integration site 6 (oncogene homolog)
• Viral integration site protein INT-6 homolog

Background

Int6 is a candidate tumor suppressor in multiple neoplasms, and in particular, breast and lung cancers. The Int6 locus was initially identified as a common insertion site (CIS) in a genetic screen for transforming sequences in a breast cancer mouse model system. Insertion of mouse mammary tumor virus (MMTV) into this locus results in the production of an amino-terminal truncated gene product. Expression of the truncated Int6 product corresponds to cellular transformation in both in vivo and in vitro systems. This gene product plays a role in regulating translation initiation and is a component of the eIF3 translation initiation complex. There is evidence that suggests that Int6 may impart a negative role in the general translational machinery while promoting an increase in the expression of a subset of stressresponsive genes. Taken together, it is of great interest to further study the mechanism by which Int6 is involved in regulating cell growth.

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: EIF3E