Mouse NEGR1 HEK293 Overexpression Lysate

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Mouse NEGR1 HEK293 Overexpression Lysate: 产品信息

产品描述
This Mouse NEGR1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of NEGR1 protein (Cat: 50601-M08H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Mouse
蛋白构建信息
A DNA sequence encoding the mouse NEGR1 (NP_001034183.1) (Met 1-Gly 318) without the pro peptide was expressed, with a C-terminal polyhistidine tag.
分子量
The secreted recombinant mouse NEGR1 comprises 298 amino acids and has a calculated molecular mass of 33 kDa. As a result of glycosylation, rmNEGR1 migrates as an approximately 47 kDa band in SDS-PAGE under reducing conditions.

Mouse NEGR1 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.

Mouse NEGR1 HEK293 Overexpression Lysate: 别称

Mouse 5330422G01Rik Overexpression Lysate; Mouse KILON Overexpression Lysate; Mouse Ntra Overexpression Lysate

NEGR1 背景信息

Neuronal Growth Regulator 1, NEGR1, also known as neurotractin, or KILON, belongs to the immunoglobulin superfamily, IgLON family. This GPI-linked cell surface glycoprotein NEGR1 is composed of three Ig-like domains and belongs to the IgLON subgroup of neural IgSF members. It is expressed in two isoforms with apparent molecular masses of 50 and 37 kD, termed L-form and S-form, respectively. NEGR1/Neurotractin participates in the regulation of neurite outgrowth in the developing brain and is expressed on the neurites of primary hippocampal neurons. Neurotractin/KILON is a trans-neural growth-promoting factor for outgrowing axons following hippocampal denervation. KILON (kindred of IgLON) and opioid-binding cell adhesion molecule, together with the limbic system-associated membrane protein and neurotrophin, belong to the IgLON subgroup of immunoglobulin superfamily. The alteration of the modulatory function of KILON/NEGR1 for the number of dendritic synapses concomitant with changes in its localization and detergent solubility during neuronal culture development. In addition to its reported role in the brain, NEGR1 is also expressed in subcutaneous adipose tissue and acts as a central 'hub' in an obesity-related transcript network.
全称
neuronal growth regulator 1
参考文献
  • Marg A, et al. (1999) Neurotractin, a novel neurite outgrowth-promoting Ig-like protein that interacts with CEPU-1 and LAMP. J Cell Biol. 145(4): 865-76.
  • Miyata S, et al. (2003) Biochemical and ultrastructural analyses of IgLON cell adhesion molecules, Kilon and OBCAM in the rat brain. Neuroscience. 117(3): 645-58.
  • Schfer M, et al. (2005) Neurotractin/kilon promotes neurite outgrowth and is expressed on reactive astrocytes after entorhinal cortex lesion. Mol Cell Neurosci. 29(4): 580-90.
  • Hashimoto T, et al. (2008) IgLON cell adhesion molecule Kilon is a crucial modulator for synapse number in hippocampal neurons. Brain Res. 1224: 1-11.
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