Human Carboxypeptidase A2 HEK293 Overexpression Lysate: 製品の情報
This Human Carboxypeptidase A2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Carboxypeptidase A2 protein (Cat: 10499-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the pre pro form of human CPA2 (NP_001859.1) (Met 1-Tyr 417) was expressed with a C-terminal polyhistidine tag.
The secreted recombinant human CPA2 existing as the proform consists of 412 amino acids and has a predicted molecular mass of 46 kDa also as estimated by SDS-PAGE under reducing conditions.
Human Carboxypeptidase A2 HEK293 Overexpression Lysate: 用法
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.
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).
安定性 & 保存条件
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 Carboxypeptidase A2 HEK293 Overexpression Lysate: 別名
Human CPA2 Overexpression Lysate
Carboxypeptidase A2 背景情報
Carboxypeptidase A2 ( CPA2 ) is a secreted pancreatic procarboxy -peptidase, and cleaves the C-terminal amide or ester bond of peptides that have a free C-terminal carboxyl group. The hydrolytic action of CPA2 was identified with a preference towards long substrates with aromatic amino acids in their C-terminal end, particularly tryptophan. CPA2 comprises a signal peptide, a pro region and a mature chain, and can be activated after cleavage of the pro peptide. Three different forms of human pancreatic procarboxypeptidase A have been isolated, and the A1 and A2 forms are always secreted as monomeric proteins with different biochemical properties.
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