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-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the human NRXN3 isoform 2 (NP_620426.2) extracellular domain (Met 1-Thr 357) was fused with the Fc region of human IgG1 at the C-terminus.
The secreted recombinant human NRXN3/Fc is a disulfide-linked homodimeric protein. The reduced monomer consists of 563 amino acids and predictes a molecular mass of 61.6 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh NRXN3/Fc monomer is approximately 75-85 kDa due to glycosylation.
Human NRXN3 HEK293 Overexpression Lysate: 使用指南
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.
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.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human NRXN3 HEK293 Overexpression Lysate: 别称
Human C14orf60 Overexpression Lysate
Neurexin-3-beta, also known as Neurexin III-beta and NRXN3, is a single-pass type I membrane protein that belongs to the neurexin family. It contains one laminin 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 has been linked to a genetic predisposition towards some conditions such as alcohol or drug addiction, or obesity.
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