Human NCAM HEK293 Overexpression Lysate: Product Information
This Human NCAM overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of NCAM protein (Cat: 10673-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human NCAM1 (NP_001070150.1) (Met1-Pro603) was expressed with a polyhistidine tag at the C-terminus.
The recombinant human NCAM1 consists 595 amino acids and predicts a molecular mass of 66.1 kDa.
Human NCAM HEK293 Overexpression Lysate: Usage Guide
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).
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human NCAM HEK293 Overexpression Lysate: Alternative Names
Human CD56 Overexpression Lysate; Human MSK39 Overexpression Lysate; Human NCAM Overexpression Lysate
NCAM Background Information
NCAM1, also known as CD56, is a neural adhesion protein (NCAM) which belongs to the immunoglobulin superfamily. NCAM is involved in neural development and in plasticity in the adult brain. UCHL1 is a novel interaction partner of both NCAM isoforms that regulates their ubiquitination and intracellular trafficking. NCAM1 is a cell adhesion molecule involved in neuron-neuron adhesion, neurite fasciculation, outgrowth of neurites, etc. NCAM1 has also been shown to be involved in the expansion of T cells and dendritic cells which play an important role in immune surveillance.
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