Contactin-6 Overexpression Lysate

Western Blot: Contactin 6 Overexpression Lysate (Adult Normal) [NBP2-10318] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Contactin 6.

• Reactivity: Hu
• Applications: WB

Contactin-6 Overexpression Lysate Summary

• Description:

  Contactin-6 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC215048
  
  Accession#: NM_014461
  
  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: CNTN6

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: 113.8 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 Contactin-6 Overexpression Lysate

• CNTN6
• contactin 6
• Contactin6
• Contactin-6
• MGC133256
• NB3

Background

The neural adhesion molecule Contactin-6, also known as NB-3, is a contactin/F3 subgroup member of immunoglobulin superfamily. It is expressed exclusively in the nervous system and mainly upregulated at the early postnatal stage during mouse brain development. Employing Northern blot analysis Kamei et al found that amongst different regions of the adult human nervous system cerebellum expressed highest level of NB-3 mRNA. The expression of NB-3 in the cerebellum increases until adulthood. In contrast, the expression in the cerebrum declines to a low level after postnatal day 7. NB-3 like other neural recognition molecules plays a vitally important role in axonal guidance during development, plasticity, and maintenance of synaptic connections in the adult brain. Cui et al recently showed that NB-3 acts as a novel Notch ligand to participate in oligodendrocyte generation. Furthermore, NB-3 triggers nuclear translocation of the Notch intracellular domain and promotes oligodendrogliogenesis from progenitor cells and differentiation of oligodendrocyte precursor cells via Deltex1. In primary oligodendrocytes, NB-3 increases myelin-associated glycoprotein transcripts. Hence, the NB-3/Notch signaling pathway may be worthwhile a closer examination for its potential for the treatment of demyelinating diseases. Human NB-3 shares with rat NB-3 86% identity in nucleotide sequences and 90% identity in amino acid sequences. FUNCTION: Contactins mediate cell surface interactions during nervous system development. Participates in oligodendrocytes generation by acting as a ligand of NOTCH1. Its association with NOTCH1 promotes NOTCH1 activation through the released notch intracellular domain (NICD) and subsequent translocation to the nucleus. Involved in motor coordination. SUBCELLULAR LOCATION: Cell membrane; lipid-anchor; GPI-anchor. ALTERNATIVE PRODUCTS: 2 named isoforms produced by alternative splicing. TISSUE SPECIFICITY: Expressed in brain. In brain, it is preferentially expressed in the accessory olfactory bulb, layers II/III and V of the cerebral cortex, piriform cortex, anterior thalamic nuclei, locus coeruleus of the pons and mesencephalic trigeminal nucleus and in Purkinje cells of the cerebellum. DEVELOPMENTAL STAGE: Highly expressed after birth, reaching a maximum at the postnatal day 7, and declines thereafter in the cerebrum, whereas it increases in the cerebellum to adulthood. MISCELLANEOUS: Mice lacking CNTN6 are viable and fertile, the formation and organization of all nuclei and layers throughout the brains are apparently normal. They are however slow to learn to stay on the rotating rod in the rotorod test during repeated trials, and display dysfunction of equilibrium and vestibular senses in the wire hang and horizontal rod-walking tests. SIMILARITY: Belongs to the immunoglobulin superfamily. Contactin family. SIMILARITY: Contains 4 fibronectin type-III domains. SIMILARITY: Contains 6 Ig-like C2-type (immunoglobulin-like) domains.

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: CNTN6