TRIAD3 Overexpression Lysate

Western Blot: TRIAD3 Overexpression Lysate (Adult Normal) [NBL1-15444] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for TRIAD3.

• Reactivity: Hu
• Applications: WB

TRIAD3 Overexpression Lysate Summary

• Description:

  TRIAD3 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC209430
  
  Accession#: NM_207116
  
  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: RNF216

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: 99.2 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 TRIAD3 Overexpression Lysate

• EC 6.3.2.-
• ring finger protein 216Ubiquitin-conjugating enzyme 7-interacting protein 1
• Triad domain-containing protein 3
• TRIAD3Zinc finger protein inhibiting NF-kappa-B
• U7I1
• UBCE7IP1ubiquitin conjugating enzyme 7 interacting protein 1
• ZINE3 ubiquitin-protein ligase RNF216

Background

This gene encodes a cytoplasmic protein which specifically colocalizes and interacts with the serine/threonine protein kinase, receptor-interacting protein (RIP). Zinc finger domains of the encoded protein are required for its interaction with RIP and for inhibition of TNF- and IL1-induced NF-kappa B activation pathways. The encoded protein may also function as an E3 ubiquitin-protein ligase which accepts ubiquitin from E2 ubiquitin-conjugating enzymes and transfers it to substrates. Several alternatively spliced transcript variants have been described for this locus but the full-length natures of only some are known. [provided by RefSeq]. Transcript Variant: This variant (2, also known as TRIAD3A) uses an alternate in-frame splice site in the 5' coding region, compared to variant 1. The resulting protein (isoform b) has a shorter N-terminus when compared to isoform a.

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: RNF216