NONO Overexpression Lysate

Western Blot: NONO Overexpression Lysate (Adult Normal) [NBP2-09748] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NONO.

• Reactivity: Hu
• Applications: WB

NONO Overexpression Lysate Summary

• Description:

  NONO Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC226579
  
  Accession#: NM_001145409
  
  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: NONO

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: 54.1 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 NONO Overexpression Lysate

• NMT5552 kDa subunit
• non-POU domain containing, octamer-binding
• NRB54non-POU-domain-containing, octamer-binding
• p54(nrb)

Background

Non-POU-domain-containing octamer binding protein (NONO) is a member of the DBHS (drosophila behavior, human splicing) domain-containing family and is an RNA- and DNA- binding protein. NONO and other DBHS domain-containing proteins are multifunctional and are reported to be involved in transcriptional regulation, mRNA processing, and DNA non-homologous end joining (NHEJ). NONO functions as a coregulator of the androgen receptor (AR) and also regulates cAMP transcriptional activity by interacting with gene promoter elements. NONO is also involved in pre-mRNA splicing through an interaction with U5 snRNA and can stimulate DNA nonhomologous end joining (NHEJ) through the interaction with ku70/G22p and ku80/XRCC5 dimers. Alternate names for NONO include 54 kDa nuclear RNA- and DNA-binding protein, p54 (nrb), 55 kDa nuclear protein, NMT55, DNA-binding p52/p100 complex, NRB54, P54, and P54NRB.

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: NONO