Human Pellino-1 Baculovirus-Insect Overexpression Lysate


Human Pellino-1 Baculovirus-Insect Overexpression Lysate: 产品信息

This Human Pellino-1 overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of Pellino-1 protein (Cat: 15180-H20B) from the overexpression lysate was verified.
Baculovirus-Insect Cells
A DNA sequence encoding the human PELI1 (NP_065702.2) (Met1-Asp418) was expressed with a N-terminal GST-tagged polyhistidine tag at the N-terminus.
The recombinant human PELI1 consists 655 amino acids and predicts a molecular mass of 74.1 kDa.

Human Pellino-1 Baculovirus-Insect Overexpression Lysate: 使用指南

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.
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.
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.
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.
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human Pellino-1 Baculovirus-Insect Overexpression Lysate: 别称

Human PRISM Overexpression Lysate

Pellino-1 背景信息

PELI is a family of E3 ubiquitin ligases that regulate protein activity through a post-translational modification, ubiquitination. PELI1 may function to control inadvertent activation of RIP3, thus preventing aberrant cell death and maintaining cellular homeostasis. PELI1 might participate in B-cell maturation or oncogenic activation of aggressive B-cell lymphomas, both during and after germinal center stages.
pellino E3 ubiquitin protein ligase 1
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