Human Cortisol Binding Globulin HEK293 Overexpression Lysate: 产品信息
This Human Cortisol Binding Globulin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Cortisol Binding Globulin protein (Cat: 10998-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human SERPINA6 (NP_001747.2) precursor (Met 1-Val 405) was fused with a polyhistidine tag at the C-terminus.
The secreted recombinant human SERPINA6 consists of 394 amino acids and predictes a molecular mass of 44 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of the protein is approximately 55-60 kDa due to glycosylation.
Human Cortisol Binding Globulin 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 Cortisol Binding Globulin HEK293 Overexpression Lysate: 别称
Human CBG Overexpression Lysate; Human SerpinA6 Overexpression Lysate
Cortisol Binding Globulin 背景信息
Corticosteroid-binding globulin (CBG), also known as SerpinA6, is a non-inhibitory member of the serine proteinase inhibitor (serpin) superfamily. It is the high-affinity transport protein for glucocorticoids in vertebrate blood. CBG is specifically cleaved by this protease at a precise site close to its carboxy-terminus. This induces a conformation change and disrupts the binding between glucocorticoids and CBG, and promotes a significant and local release of glucocorticoids (over 90% of them are bound to CBG in human plasma). In this context, CBG directs glucocorticoids to sites of inflammation, and plays in consequence a crucial role in efficient glucocorticoid action in physiology. The SerpinA6 protein is mainly secreted by the liver. This negative acute phase protein regulates free cortisol levels in the blood and distributes cortisol to its target tissues. SerpinA6 deficiency is an extremely rare hereditary disorder characterized by reduced corticosteroid-binding capacity with normal or low plasma corticosteroid-binding globulin concentration, and normal or low basal cortisol levels associated with hypo-/hypertension and muscle fatigue. There are three heritable, human CBG gene mutations that can reduce CBG-cortisol binding affinity and/or reduce circulating CBG levels.
serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 6
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