This Human SerpinA3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of SerpinA3 protein (Cat: 10307-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human SERPINA3 (NP_001076.2) (Met 1-Ala 423) was expressed, with a C-terminal polyhistidine tag.
The recombinant human SERPINA3 consisting of 409 amino acids and has a calculated molecular mass of 46.5 kDa. As a result of different glycosylation, the apparent molecular mass of the recombinant protein is approximately 45 kDa and 55-70 kDa in SDS-PAGE under reducing conditions.
Human SerpinA3 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 SerpinA3 HEK293 Overexpression Lysate: 别称
Human AACT Overexpression Lysate; Human ACT Overexpression Lysate; Human GIG24 Overexpression Lysate; Human GIG25 Overexpression Lysate; Human MGC88254 Overexpression Lysate; Human SERPINA3 Overexpression Lysate
SerpinA3, also known as Alpha 1-antichymotrypsin (AACT), is a plasma alpha globulin glycoprotein, and is a member of serpin superfamily of the serine protease inhibitors consisting of at least 35 members. SerpinA3 has been demonstrated to inhibit the activity of certain serine proteases, such as cathepsin G found in neutrophils, and chymases present in mast cells, by inducing a major conformational rearrangement, and thus protects some tissues from damage caused by proteolytic enzymes. This enzyme is produced primarily in the liver, and is identified as an acute-phase inflammatory protein. SerpinA3 deficiency has been associated with liver disease, and mutations of this gene have been observed in patients with Parkinson disease and chronic obstructive pulmonary disease. Besides, ACT gene polymorphism has been implicated with Alzheimer’s disease (AD), cerebral amyloid angiopathy (CAA), as well as stroke, since SerpinA3 is a major constituent of the plaques in AD and an inhibitor of amyloid beta peptide degradation.
serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 3
Nilsson, L.N. et al., 2001, J. Neurosci. 21: 1444-1451.
Ikari, Y. et al., 2001, J. Biol. Chem. 276: 11798-11803.
Vila, N. et al., 2000, Stroke. 31: 2103-2105
Eriksson, S. et al., 1995, Proc. Natl. Acad. Sci. USA. 92: 2313-2317.
Skeel, A. et al., 2001, J. Biol. Chem. 276: 21932-21937.