SARS-CoV-2 (2019-nCoV) |新冠病毒 Spike HEK293 Cell Lysate (WB positive control): 产品信息
产品描述：This CoV spike overexpression lysate was created in Human Cells and intented for use as a Western blot (WB) positive control. Purification of CoV spike protein (Cat: 40591-V05H1) from the overexpression lysate was verified.
表达宿主：Human Cells
蛋白构建信息：A DNA sequence encoding the 2019-nCoV spike protein S1 was expressed with the Fc region of mouse IgG1 at the C-terminus.
分子量：The recombinant 2019-nCoV spike protein S1 consists of 904 amino acids and predicts a molecular mass of 101.4 kDa.
2019-nCoV Spike HEK293 Cell Lysate (WB positive control): 使用指南
制备方法
Cell lysate was prepared by homogenization 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. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
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.
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