Human Neurofascin HEK293 Overexpression Lysate

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Human Neurofascin HEK293 Overexpression Lysate: 产品信息

产品描述
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.
表达宿主
HEK293 Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human NFASC (XP_011507630.1) (Met1-Gln939) was expressed with the Fc region of human IgG1 at the C-terminus.
分子量
The recombinant human NFASC consists 1156 amino acids and predicts a molecular mass of 130 kDa.

Human Neurofascin HEK293 Overexpression Lysate: 使用指南

制备方法
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.
裂解缓冲液
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.
使用建议
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.
缓冲液
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.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human Neurofascin HEK293 Overexpression Lysate: 别称

Human NF Overexpression Lysate; Human NRCAML Overexpression Lysate

Neurofascin 背景信息

NFASC (Neurofascin, also known as NRCAML, NEDCPMD, and NF) is a Protein Coding gene. 13 alternatively spliced human isoforms have been reported. NFASC 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. The protein functions in neurite outgrowth, neurite fasciculation, and organization of the axon initial segment (AIS) and nodes of Ranvier on axons during early development. NFASC links the AIS extracellular matrix to the intracellular cytoskeleton. It is broadly expressed in the brain, kidney, and other tissues. Diseases associated with NFASC include Neurodevelopmental Disorder With Central And Peripheral Motor Dysfunction and Demyelinating Polyneuropathy.
全称
neurofascin
参考文献
  • 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.
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