Epsin-2 Overexpression Lysate

Western Blot: Epsin-2 Overexpression Lysate (Adult Normal) [NBP2-05424] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Epsin-2.

• Reactivity: Hu
• Applications: WB

Epsin-2 Overexpression Lysate Summary

• Description:

  Epsin-2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC210899
  
  Accession#: NM_148921
  
  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: EPN2

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: 62.1 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 Epsin-2 Overexpression Lysate

• Eps15 binding protein
• EPS-15-interacting protein 2
• epsin 2
• epsin-2
• KIAA1065EHB21

Background

Elucidation of the mechanism by which receptor tyrosine kinases (RTKs) modulate cellular physiology in response to stimuli is critical to the understanding of growth regulation. Miscues in RTK signaling pathways can result in cellular transformation and ultimately in cancer. Two novel EGF receptor substrates designated EGF-receptor pathway substrates 8 and 15, or Eps8 and Eps15, have been described. Epsin is a 90 kDa binding partner to Eps15. Both epsin and Eps15 have an ubiquitous tissue distribution but are concentrated in presynaptic nerve terminals specialized for the Clathrin-mediated endocytosis of synaptic vesicles. Disruption of epsin function blocks Clathrinmediated endocytosis. Epsin along with its binding partner Eps15 is proposed to be involved in the assistance of Clathrin coat rearrangement during Clathrin coated pit invagination. Epsin 2 and epsin 2a are also associated with Clathrin-mediated endocytosis and are enriched in the brain in the peri-Golgi region.

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: EPN2