Human Cbl-c Baculovirus-Insect cells Overexpression Lysate

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Human Cbl-c Baculovirus-Insect cells Overexpression Lysate: 产品信息

产品描述
This Human Cbl-c overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Cbl-c protein (Cat: 15659-H20B) from the overexpression lysate was verified.
表达宿主
Baculovirus-Insect cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human CBLC (AAH28915.1) (Met1-Ala474) was expressed with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
分子量
The recombinant human CBLC chimera consists of 711 amino acids and has a calculated molecular mass of 80.2 kDa. The recombinant protein migrates approximately 66 kDa band in SDS-PAGE under reducing conditions.

Human Cbl-c 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.

Human Cbl-c Baculovirus-Insect cells Overexpression Lysate: 别称

Human CBL-3 Overexpression Lysate; Human CBL-SL Overexpression Lysate; Human RNF57 Overexpression Lysate

Cbl-c 背景信息

CBL proteins, such as Cbl-c, are phosphorylated upon activation of a variety of receptors that signal via protein tyrosine kinases. Through interactions with proteins containing SRC homology-2 (SH2) and SH3 domains, CBL proteins modulate downstream cell signaling. Cbl-c is a member of the Cbl family of E3 ubiquitin ligases. Expression of Cbl-c gene may be restricted to epithelial cells, and alternatively spliced transcript variants encoding multiple isoforms have been observed for Cbl-c gene.
全称
Cbl proto-oncogene C, E3 ubiquitin protein ligase
参考文献
  • Kim M. et al., 1999, Gene. 239 (1): 145-54.
  • Courbard JR. et al., 2003, J. Biol. Chem. 277 (47): 45267-75.
  • Keane MM. et al., 1999, Oncogene. 18 (22): 3365-75.
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