Mouse EphB1 HEK293 Overexpression Lysate

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Mouse EphB1 HEK293 Overexpression Lysate: 製品の情報

製品の説明
This Mouse EphB1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of EphB1 protein (Cat: 50479-M08H) from the overexpression lysate was verified.
発現ホスト
HEK293 Cells
Mouse
タンパク質構築情報
A DNA sequence encoding the mouse EPHB1 (Q8CBF3-1) extracellular domain (Met 1-Leu 539) was expressed, with a polyhistidine tag at the C-terminus.
分子量
The secreted recombinant mouse EPHB1 consists of 533 amino acids and has a predicted molecular mass of 59.6 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rm EPHB1 is approximately 64 kDa due to glycosylation.

Mouse EphB1 HEK293 Overexpression Lysate: 用法

調製方法
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.
溶解バッファー
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.
おすすめの用法
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.
バッファー
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
安定性 & 保存条件
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.
アプリケーション
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Mouse EphB1 HEK293 Overexpression Lysate: 別名

Mouse 9330129L11 Overexpression Lysate; Mouse AW488255 Overexpression Lysate; Mouse C130099E04Rik Overexpression Lysate; Mouse Cek6 Overexpression Lysate; Mouse Elk Overexpression Lysate; Mouse Elkh Overexpression Lysate; Mouse ENSMUSG00000074119 Overexpression Lysate; Mouse Hek6 Overexpression Lysate; Mouse Net Overexpression Lysate

EphB1 背景情報

Ephrin type-B receptor 1, also known as EphB1, belongs to the ephrin receptor subfamily of the protein-tyrosine kinase family which 16 known receptors (14 found in mammals) are involved: EPHA1, EPHA2, EPHA3, EPHA4, EPHA5, EPHA6, EPHA7, EPHA8, EPHA9, EPHA1, EPHB1, EPHB2, EPHB3, EPHB4, EPHB5, EPHB6. EphB2 receptor tyrosine kinase phosphorylates syndecan-2 and that this phosphorylation event is crucial for syndecan-2 clustering and spine formation. The Eph family of receptor tyrosine kinases (comprising EphA and EphB receptors) has been implicated in synapse formation and the regulation of synaptic function and plasticity6. Ephrin receptors are components of cell signalling pathways involved in animal growth and development, forming the largest sub-family of receptor tyrosine kinases (RTKs). Ligand-mediated activation of Ephs induce various important downstream effects and Eph receptors have been studied for their potential roles in the development of cancer. EphB receptor tyrosine kinases are enriched at synapses, suggesting that these receptors play a role in synapse formation or function. We find that EphrinB binding to EphB induces a direct interaction of EphB with NMDA-type glutamate receptors. This interaction occurs at the cell surface and is mediated by the extracellular regions of the two receptors, but does not require the kinase activity of EphB.
完全な名称
EPH receptor B1
参考文献
  • Dalva MB, et al. (2000) EphB receptors interact with NMDA receptors and regulate excitatory synapse formation. Cell. 103(6): 945-56.
  • Takasu MA, et al. (2002) Modulation of NMDA receptor-dependent calcium influx and gene expression through EphB receptors. Science. 295(5554): 491-5.
  • Adams RH, et al. (1999) Roles of ephrinB ligands and EphB receptors in cardiovascular development: demarcation of arterial/venous domains, vascular morphogenesis, and sprouting angiogenesis. Genes Dev. 13(3): 295-306.
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