Mouse Smad5 Baculovirus-Insect cells Overexpression Lysate

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Mouse Smad5 Baculovirus-Insect cells Overexpression Lysate: 产品信息

产品描述
This Mouse Smad5 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Smad5 protein (Cat: 50728-M20B) from the overexpression lysate was verified.
表达宿主
Baculovirus-Insect cells
种属
Mouse
蛋白构建信息
A DNA sequence encoding the mouse SMAD5 (P97454) (Thr2-Ser465) was expressed with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
分子量
The recombinant mouse SMAD5/GST chimera consists of 701 amino acids and has a calculated molecular mass of 79.9 kDa. The recombinant protein migrates as an approximately 75-88 kDa band in SDS-PAGE under reducing conditions.

Mouse Smad5 Baculovirus-Insect cells 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 Smad5 Baculovirus-Insect cells Overexpression Lysate: 别称

Mouse 1110051M15Rik Overexpression Lysate; Mouse AI451355 Overexpression Lysate; Mouse Dwf-C Overexpression Lysate; Mouse Madh5 Overexpression Lysate; Mouse MusMLP Overexpression Lysate

Smad5 背景信息

SMAD5 is a member of the SMAD family. Members of this family mediate signal transduction by the TGF-beta/activin/BMP-2/4 cytokine superfamily from receptor Ser/Thr protein kinases at the cell surface to the nucleus. SMAD5 is involved in the TGF-beta signaling pathway that results in an inhibition of the proliferation of hematopoietic progenitor cells. It is also involved in cell signalling and modulates signals of bone morphogenetic proteins (BMP's). The binding of ligands causes the oligomerization and phosphorylation of the SMAD5 protein. SMAD5 is a receptor regulated SMAD (R-SMAD) and is activated by bone morphogenetic protein type 1 receptor kinase.
全称
SMAD family member 5
Related Pathways
  • TGF-beta Signaling
    TGF-beta Signaling
参考文献
  • Vinayagam A. et al., 2011, Sci Signal. 4 (189): rs8.
  • Sangadala S. et al., 2007, J Biomol Struct Dyn. 25 (1): 11-23.
  • Riggins GJ. et al., 1996, Nat Genet. 13 (3): 347-9.
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