Human NRXN3 HEK293 Overexpression Lysate

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Human NRXN3 HEK293 Overexpression Lysate: Product Information

Product Description
This Human NRXN3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of NRXN3 protein (Cat: 11843-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human NRXN3 beta isoform 2 (NP_620426.2) extracellular domain (Met 1-Thr 357) was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human NRXN3 consists of 333 amino acids and predictes a molecular mass of 36 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhNRXN3 is approximately 50-60 kDa due to glycosylation.

Human NRXN3 HEK293 Overexpression Lysate: Usage Guide

Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human NRXN3 HEK293 Overexpression Lysate: Alternative Names

Human C14orf60 Overexpression Lysate

NRXN3 Background Information

Neurexin-3-beta, also known as Neurexin III-beta and NRXN3, is a single-pass type I membrane protein which belongs to theneurexin family. It contains onelaminin G-like domain. It is a neuronal cell surface protein that may be involved in cell recognition and cell adhesion. Neurexins are a family of proteins that function in the vertebrate nervous system as cell adhesion molecules and receptors. They are encoded by several unlinked genes of which two, NRXN1 and NRXN3, are among the largest known human genes. Three of the genes ( NRXN1, NRXN2, NRXN3 ) utilize two alternate promoters and include numerous alternatively spliced exons to generate thousands of distinct mRNA transcripts and protein isoforms. The majority of transcripts are produced from the upstream promoter and encode alpha-neurexin isoforms; a much smaller number of transcripts are produced from the downstream promoter and encode beta-neurexin isoforms. The alpha-neurexins contain EGF-like sequences and laminin G domains, and have been shown to interact with neurexophilins. The beta-neurexins lack EGF-like sequences and contain fewer laminin G domains than alpha-neurexins. NRXN3 have been linked to genetic predisposition towards a number of conditions such as alcohol or drug addiction, or obesity.
Full Name
neurexin 3
References
  • Occhi G. et al., 2002, Biochem. Biophys. Res. Commun. 298:151-5.
  • Rowen L, et al., 2002, Genomics. 79 (4): 587-97.
  • Olsen J.V. et al., 2006, Cell 127:635-48.
  • Hishimoto A, et al., 2007, Human Molecular Genetics.16 (23): 2880-91.
  • Oppermann F.S. et al., 2009, Mol. Cell. Proteomics 8:1751-64. 
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