Neosartorya fumigata AspF9 HEK293 Overexpression Lysate

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Neosartorya fumigata AspF9 HEK293 Overexpression Lysate: 产品信息

产品描述
This Neosartorya fumigata AspF9 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of AspF9 protein (Cat: 69038-A08H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Neosartorya fumigata
蛋白构建信息
A DNA sequence encoding the neosartorya fumigata AspF9 (XP_752985.1) (Met1-Gly370) was expressed with a polyhistidine tag at the C-terminus.
分子量
The recombinant neosartorya fumigata AspF9 consists of 362 amino acids and predicts a molecular mass of 37.5 kDa.

Neosartorya fumigata AspF9 HEK293 Overexpression Lysate: 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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