Mouse Pleiotrophin/PTN HEK293 Overexpression Lysate

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Mouse Pleiotrophin/PTN HEK293 Overexpression Lysate: 产品信息

产品描述
This Mouse Pleiotrophin/PTN overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Pleiotrophin/PTN protein (Cat: 51000-M01H) 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 PTN (P63089) (Gly 33-Asp 168) was fused with the Fc region of human IgG1 at the N-terminus.
分子量
The recombinant mouse PTN/Fc is a disulfide-linked homodimer. The reduced monomer comprises 396 amino acids and has a calculated molecular mass of 43.7 kDa. The apparent molecular mass of the monomer is approximately 47 kDa in SDS-PAGE under reducing conditions.

Mouse Pleiotrophin/PTN 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 Pleiotrophin/PTN HEK293 Overexpression Lysate: 别称

Mouse HARP Overexpression Lysate; Mouse HB-GAM Overexpression Lysate; Mouse HBBN Overexpression Lysate; Mouse HBGF-8 Overexpression Lysate; Mouse HBNF Overexpression Lysate; Mouse OSF Overexpression Lysate; Mouse Osf-1 Overexpression Lysate; Mouse Osf1 Overexpression Lysate

Pleiotrophin/PTN 背景信息

HB-GAM belongs to the pleiotrophin family. During embryonic and early postnatal development, HB-GAM is expressed in the central and peripheral nervous system and also in several non-neural tissues, notably lung, kidney, gut and bone. While in the adult central nervous system, it is expressed in an activity-dependent manner in the hippocampus where it can suppress long term potentiation induction. HB-GAM has a low expression in other areas of the adult brain, but it can be induced by ischemic insults, or targeted neuronal damage in the entorhinal cortex or in the substantia nigra pars compacta. It is structurally related to midkine and retinoic acid induced heparin-binding protein and has a high affinity for heparin. HB-GAM binds anaplastic lymphoma kinase (ALK) which induces MAPK pathway activation, an important step in the anti-apoptotic signaling of PTN and regulation of cell proliferation. It also functions as a secreted growth factor and induces neurite outgrowth and which is mitogenic for fibroblasts, epithelial, and endothelial cells.
全称
pleiotrophin
参考文献
  • Vanderwinden JM, et al. (1992) Cellular distribution of the new growth factor pleiotrophin (HB-GAM) mRNA in developing and adult rat tissues. Anat Embryol. 186(4):387-406.
  • Lauri SE, et al. (1996) Activity-induced enhancement of HB-GAM expression in rat hippocampal slices. Neuroreport. 7(10):1670-4.
  • Pavlov I, et al. (2002) Role of heparin-binding growth-associated molecule (HB-GAM) in hippocampal LTP and spatial learning revealed by studies on overexpressing and knockout mice. Mol Cell Neurosci. 20(2):330-42.
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