EPS15R Overexpression Lysate

Western Blot: EPS15R Overexpression Lysate (Adult Normal) [NBL1-10302] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for EPS15R.

• Reactivity: Hu
• Applications: WB

EPS15R Overexpression Lysate Summary

• Description:

  EPS15R Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC215194
  
  Accession#: NM_021235
  
  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: EPS15L1

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: 94.1 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 EPS15R Overexpression Lysate

• epidermal growth factor receptor pathway substrate 15-like 1
• epidermal growth factor receptor substrate 15-like 1
• epidermal growth factor receptor substrate EPS15R
• EPS15R
• Eps15-related protein

Background

EPS15 and EPS15R are substrates of the epidermal growth factor (EGF) receptor kinase that are characterized by the presence of a protein:protein interaction domain, the EH domain, and by their ability to bind to the clathrin adaptor protein complex adaptor protein 2. Evidence suggests that eps15 and eps15R are involved in endocytosis (1). The most striking difference between these two related proteins is that Eps15R is also found in the nucleus, whereas Eps15 is excluded from this compartment at steady state (2). Eps15R plays a key role in clathrin-mediated endocytosis of transmembrane receptors. Using a mutational approach, it has been found that the second ubiquitin-interacting motif (UIM) of Eps15 and Eps15R is essential for their ubiquitination (3)

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: EPS15L1