NLE1 Overexpression Lysate

Western Blot: NLE1 Overexpression Lysate (Adult Normal) [NBL1-13664] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NLE1.

• Reactivity: Hu
• Applications: WB

NLE1 Overexpression Lysate Summary

• Description:

  NLE1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC204114
  
  Accession#: NM_018096
  
  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: NLE1

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: 53.1 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 NLE1 Overexpression Lysate

• FLJ10458
• Nle
• Notchless gene homolog
• notchless homolog 1 (Drosophila)
• notchless protein homolog 1

Background

The Notch signaling pathway is an evolutionary conserved system that is involved in intracellular communication. Notch receptors play an important role in development and cell-fate decisions. Notchless is a loss-of-function mutant allele that encodes for protein NLE1 (notchless homolog 1). NLE1 is a 485 amino acid WD40-repeat protein that binds to the cytoplasmic domain of Notch, regulating its signaling activity in Drosophila melanogaster and in mice. Deletion of the NLE1 gene in mice during the early stages of development results in embryonic death, while gene deletion in the late stages of development leads to activation of a caspase-3-dependent apoptotic pathway.In plants, NLE1 is crucial for normal cellular growth and development. Underexpression during shoot proliferation causes pleiotropic defects such as delayed flowering and abnormal organ maturation. It may also play a role in 60S ribosomal subunit biogenesis in yeast. NLE1 contains eight WD40 domains and produces one isoform due to alternative splicing.

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: NLE1