DEDD2 Overexpression Lysate

Western Blot: DEDD2 Overexpression Lysate (Adult Normal) [NBL1-09816] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for DEDD2.

• Reactivity: Hu
• Applications: WB

DEDD2 Overexpression Lysate Summary

• Description:

  DEDD2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC206798
  
  Accession#: NM_133328
  
  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: DEDD2

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: 36 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 DEDD2 Overexpression Lysate

• death effector domain containing 2
• death effector domain-containing DNA binding protein 2
• DED-containing protein FLAME-3
• DNA-binding death effector domain-containing protein 2
• FADD-like apoptotic molecule 3
• FLAME3
• FLAME-3

Background

Apoptosis and inflammation are important cellular processes that are highly regulated through specific protein-protein interactions (PPI). Proteins involved in these signaling cascades often carry PPI domains belonging to the death-domain superfamily (reviewed in Kohl A and MG Grutter, 2004). These PPI domains include the death effector domain (DED), the death domain (DD), the caspase activation and recruitment domain (CARD) and the pyrin domain (PYD). PPI domains are structurally related and comprised of a six alpha helical structure; within proteins PPI domains can be found in isolation or in combination with other domains. DEDD2/Flame-3 (DED containing DNA binding2) belongs to a family of single DED-containing proteins that is targeted to the nucleus (reviewed in Barnhart et al, 2003; Slvibst ry sl, 2003). Several DED-containing proteins are involved in the regulation of apoptosis through their interactions with DED-containing caspases. It is thought that DEDD2 and its closely related homologue DEDD may mediate cell death by binding to the DED-containing caspases -8 and -10, and targeting them to the nucleus following death receptor induced apoptosis. However, the function of DEDD2 and DEDD remain to be fully elucidated, and they may also have roles in cellular processes other than apoptosis. This antibody recognizes DEDD2/Flame-3; human DED2/Flame-3 is a 326 amino acid protein.

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: DEDD2