Human Neurofascin HEK293 Overexpression Lysate 产品信息
Product Description
This Human Neurofascin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Neurofascin protein (Cat: 15694-H02H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human NFASC (NP_001005388.2) (Met1-Gln939) was expressed with the Fc region of human IgG1 at the C-terminus.
Molecule Mass
The recombinant human NFASC consists 1156 amino acids and predicts a molecular mass of 130 kDa.
Human Neurofascin 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 Neurofascin HEK293 Overexpression Lysate Alternative Names
Human NF Overexpression Lysate;Human NRCAML Overexpression Lysate
Neurofascin Background Information
NFASC, also known as neurofascin, belongs to the immunoglobulin superfamily, L1/neurofascin/NgCAM family. It contains 5 fibronectin type-III domains and 6 Ig-like C2-type (immunoglobulin-like) domains. NFASC functions in neurite outgrowth, neurite fasciculation, and organization of the axon initial segment (AIS) and nodes of Ranvier on axons during early development. Both the AIS and nodes of Ranvier contain high densities of voltage-gated Na+ (Nav) channels which are clustered by interactions with cytoskeletal and scaffolding proteins including this protein, gliomedin, ankyrin 3 (ankyrin-G), and betaIV spectrin. NFASC links the AIS extracellular matrix to the intracellular cytoskeleton.
References
Volkmer H. et al., 1992, J Cell Biol. 118 (1): 149-61. Burmeister M. et al., 1996, Mamm Genome. 7 (7): 558-9. Ango F. et al., 2004, Cell. 119 (2): 257-72.