Human Carboxypeptidase A HEK293 Overexpression Lysate: Product Information
This Human Carboxypeptidase A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Carboxypeptidase A protein (Cat: 10504-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human CPA1 precursor (NP_001859.1) (Met 1-Tyr 419) was expressed with a C-terminal polyhistidine tag.
The secreted recombinant human CPA1 (pro form) consists of 414 amino acids and has a predicted molecular mass of 47 kDa. In SDS-PAGE under reducing conditions, it migrates with the apparent molecular mass of 43 kDa.
Human Carboxypeptidase A HEK293 Overexpression Lysate: Usage Guide
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).
Stability & Storage
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 A HEK293 Overexpression Lysate: Alternative Names
Human CPA Overexpression Lysate
Carboxypeptidase A Background Information
Carboxypeptidase A1 （CPA1）is secreted as a pancreatic procarboxypeptidase, and cleaves the C-terminal amide or ester bond of peptides that have a free C-terminal carboxyl group, with the preference of residues with aromatic or branched aliphatic side chains. CPA1 comprises a signal peptide, a pro region and a mature chain, and can be activated after cleavage of the pro peptide. In contrast to procarboxypeptidase B which was always secreted by the pancreas as a monomer, procarboxypeptidase A occurs as a monomer and/or associated to one or two functionally different proteins, such as zymogen E, and is involved in zymogen inhibition. Three different forms of human pancreatic procarboxypeptidase A have been isolated.
carboxypeptidase A1 (pancreatic)
Catasus, L. et al., 1992, Biochem. J. 287: 299-303.
Moulard, M. et al., 1990, FEBS. Lett. 261: 179-183.
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