Human R-Cadherin/CDH4 HEK293 Overexpression Lysate

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Human R-Cadherin/CDH4 HEK293 Overexpression Lysate: 製品の情報

製品の説明
This Human R-Cadherin/CDH4 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of R-Cadherin/CDH4 protein (Cat: 10230-H08H) from the overexpression lysate was verified.
発現ホスト
HEK293 Cells
Human
タンパク質構築情報
A DNA sequence encoding the extracellular domain of human CAD4 (NP_001785.2) (Met 1-Ala 734) was expressed with a fused polyhistidine tag at the C-terminus.
分子量
The recombinant pro form of human CAD4 comprises 725 amino acids and predicts a molecular mass of 80 kDa. As a result of glycosylation, rhCAD4 migrates as an approximately 90-100 kDa protein in SDS-PAGE under reducing conditions.

Human R-Cadherin/CDH4 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.

Human R-Cadherin/CDH4 HEK293 Overexpression Lysate: 別名

Human CAD4 Overexpression Lysate; Human R-CAD Overexpression Lysate; Human RCAD Overexpression Lysate

R-Cadherin/CDH4 背景情報

The cadherin superfamily is a large family that engage in both homo- and heterotypic, calcium-dependent, cell-cell adhesion events, and can be divided into at least four subfamilies based on the extracellular (EC) regions and cytoplasmic domains, that is: classical cadherins, desmosomal cadherins, protocadherins, and cadherin-like molecules. Human cadherin 4, type 1, R-cadherin (retinal), also known as CDH4, CAD4 and RCAD, is a classical cadherin from the cadherin superfamily. It is a calcium-dependent adhesion molecule and a type I transmembrane glycoprotein composed of five extracellular cadherin repeats, a transmembrane region and a highly conserved cytoplasmic tail. CDH4 is thought to play an important role during brain segmentation and neuronal outgrowth, and also exerts critical actions in kidney and muscle development. CDH4 is expressed in vascular smooth muscle, pancreatic β-cells, thyroid follicular cells, sensory neurons of the dorsal root ganglia, and, possibly, astrocytes and endothelium of the retina. As a classic cadherin, CDH4 forms both homodimers and heterodimers with N-cadherin. The extracellular region of human CDH4 is 96% aa identical to that of mouse CDH4.
完全な名称
cadherin 4, type 1, R-cadherin (retinal)
参考文献
  • Suzuki, S. et al., 1991, Cell Regul. 2: 261-270.
  • Matsunami, H. et al., 1993, J. Cell Sci. 106: 401-409.
  • Tanihara, H. et al.,1995, Cell Adhes. Commun. 2 (1): 15-26.
  • Shan, W-S., et al., 2000, J. Cell Biol. 148: 579-590.
  • Fagman, H. et al, 2003, Endocrinology. 144: 3618-3624.
  • Koch, A.W. et al., 2004, Cell. Mol. Life Sci. 61: 1884-1895.
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