Human ABP1/AOC1 HEK293 Overexpression Lysate: 产品信息
This Human ABP1/AOC1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ABP1/AOC1 protein (Cat: 13623-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human AOC1 (AAH14093.1) (Met1-Val751) was expressed with a polyhistidine tag at the C-terminus.
The recombinant human AOC1 consists 743 amino acids and predicts a molecular mass of 84.9 kDa.
Human ABP1/AOC1 HEK293 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.
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 ABP1/AOC1 HEK293 Overexpression Lysate: 别称
Human ABP Overexpression Lysate; Human ABP1 Overexpression Lysate; Human DAO Overexpression Lysate; Human DAO1 Overexpression Lysate; Human KAO Overexpression Lysate
Amine oxidase copper-containing 1 (AOC1; formerly known as amiloride-binding protein 1) is a secreted glycoprotein that catalyzes the degradation of putrescine and histamine. Polyamines and their diamine precursor putrescine are ubiquitous to all organisms and fulfill pivotal functions in cell growth and proliferation. That the Wilms tumor protein, WT1, which is necessary for normal kidney development, activates transcription of the AOC1 gene. Expression of a firefly luciferase reporter under control of the proximal AOC1 promoter was significantly enhanced by co-transfection of a WT1 expression construct. Binding of WT1 protein to a cis-regulatory element in the AOC1 promoter was confirmed by electrophoretic mobility shift assay and chromatin immunoprecipitation. WT1-dependent control of polyamine breakdown, which is mediated by changes in AOC1 expression, has a role in kidney organogenesis.