RNF25 Overexpression Lysate

Western Blot: RNF25 Overexpression Lysate (Adult Normal) [NBP2-06433] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for RNF25.

• Reactivity: Hu
• Applications: WB

RNF25 Overexpression Lysate Summary

• Description:

  RNF25 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC204829
  
  Accession#: NM_022453
  
  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: RNF25

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: 51 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 RNF25 Overexpression Lysate

• AO7
• EC 6.3.2.-
• FLJ13906
• ring finger protein 25E3 ubiquitin-protein ligase RNF25
• ring finger protein AO7

Background

RING finger protein 25 (RNF25, also named AO7) contains a RING finger domain and is ubiquitously expressed in various tissues. RNF25 was initially identified in a yeast two-hybrid screen of a murine T-cell library by using UbcH5b, an E2 enzyme, as bait. RNF25 has also been shown to act as a putative E3 ligase, at least in vitro. RNF25 localizes predominantly in the nucleus and supports the transcriptional activity of NF-kB by interacting with p65 in vivo upon stimulation with TNF. Yeast two-hybrid data also suggest that RNF25 interacts with a number of other molecules which may be potential ubiquitin ligase substrates. Among these are molecules that have critical roles in signal transduction and in regulation of translation (personal communication, Allan Weissman, CCR-NCI, Bethesda, MD).

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: RNF25