SMURF2 Overexpression Lysate

Western Blot: SMURF 2 Overexpression Lysate (Adult Normal) [NBL1-16260] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for SMURF 2.

• Reactivity: Hu
• Applications: WB

SMURF2 Overexpression Lysate Summary

• Description:

  SMURF2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC210866
  
  Accession#: NM_022739
  
  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: SMURF2

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: 86 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 SMURF2 Overexpression Lysate

• DKFZp686F0270
• E3 ubiquitin ligase SMURF2
• E3 ubiquitin-protein ligase SMURF2
• EC 6.3.2
• EC 6.3.2.-
• hSMURF2
• MGC138150
• SMAD specific E3 ubiquitin protein ligase 2
• SMAD ubiquitination regulatory factor 2
• SMAD-specific E3 ubiquitin-protein ligase 2
• SMURF2

Background

Smad ubiquitination regulatory factor proteins (Smurf1 and Smurf2) are E3 ubiquitin ligase that belongs to the Hect family. Smurf proteins play an important role as regulators in TGF-beta pathway by ubiquitinating Smads and Smads associated proteins for proteasome degradation (1). Specifically, Smurf1 interacts with Smad1 and Smad5 for degradation, while Smurf2 ubiquitinates Smad1 and Smad2. Smads also functions to recruit Smurfs to various pathway components such as TGF-beta and SnoN. In particular, Smad7 acts as an adaptor protein between Smurfs and TGF-Beta receptors, allowing the receptors to be marked by Smurfs for degradation (2). Smurf2 interacts with all members of Smad family except for Smad4 and it is expressed various tissues and cell lines, such as placenta and ovarian cancer cell lines. Smurf2 has been implicated in the tumor formation and diseases progression (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: SMURF2