Human CDK1 & CCNE1 Heterodimer Baculovirus-Insect cells Overexpression Lysate

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Human CDK1 & CCNE1 Heterodimer Baculovirus-Insect cells Overexpression Lysate: 产品信息

产品描述
This Human CDK1 & CCNE1 Heterodimer overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of CDK1 & CCNE1 Heterodimer protein (Cat: CT027-H0907B) 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 CDC2 (NP_001777.1) (Met1-Met297) was expressed with a GST tag at the N-terminus, constructed the plasmid 1; A DNA sequence encoding the human CCNE1 (NP_001229.1) (Met1-Ala410) was expressed with a polyhistidine tag at the N-terminus, constructed the plasmid 2. The two plasmids were co-expressed and the heterotrimer was purified.
分子量
The recombinant heterotrimer of human CDC2 & CCNE1 comprises 950 (522+428) amino acids and has a calculated molecular mass of 109.7 (60.4+49.3) kDa.

Human CDK1 & CCNE1 Heterodimer 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 CDK1 & CCNE1 Heterodimer Baculovirus-Insect cells Overexpression Lysate: 别称

Human CCNE Overexpression Lysate; Human CDC2 Overexpression Lysate; Human CDC28A Overexpression Lysate; Human CDK1 Overexpression Lysate; Human CDKN1 Overexpression Lysate; Human P34CDC2 & CCNE1 Overexpression Lysate
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