Human TBG HEK293 Overexpression Lysate

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Human TBG HEK293 Overexpression Lysate: Product Information

Product Description
This Human TBG overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TBG protein (Cat: 10993-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human SERPINA7 (NP_000345.2) (Met 1-Ala 415) was expressed, fused with a polyhistidine tag at the C-terminus.
Molecule Mass
The secreted recombinant human SERPINA7 comprises 406 amino acids and has a predicted molecular mass of 45.5 kDa. As a result of glycosylation, it migrates as an approximately 55-60 kDa band in SDS-PAGE under reducing conditions.

Human TBG HEK293 Overexpression Lysate: Usage Guide

Preparation Method
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.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
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.
Sample Buffer
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human TBG HEK293 Overexpression Lysate: Alternative Names

Human Serpin A7 Overexpression Lysate; Human TBG Overexpression Lysate; Human Thyroxine Binding Globulin Overexpression Lysate

TBG Background Information

Thyroxine-binding globulin, also known as T4-binding globulin, Serpin A7 and TBG, is a secreted protein which belongs to theserpin family. TBG is synthesized primarily in theliveras a 54 kDa protein.TBG is genomically aserpin, although it has no inhibitory function like many other members of this class of proteins. TBG binds thyroid hormone in circulation. It is one of three proteins(along with transthyretin and albumin) responsible for carrying thethyroidhormones thyroxine(T4) and 3,5,3’-triiodothyronine (T3) in the bloodstream. Of these three proteins, TBG has the highest affinity for T4 and T3, but is present in the lowest concentration. Despite its low concentration, TBG carries the majority of T4 inserum. Due to the very low serum concentration of T4 and T3, TBG is rarely more than 25% saturated with itsligand. Unlike transthyretin and albumin, TBG has a single binding site for T4/T3. TBG tests are sometimes used in finding the reason for elevated or diminished levels of thyroid hormone.
Full Name
serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 7
References
  • Flink I.L., et al.,(1986), Complete amino acid sequence of human thyroxine-binding globulin deduced from cloned DNA: close homology to the serine antiproteases. Proc. Natl. Acad. Sci. U.S.A. 83:7708-7712.
  • Akbari M.T., et al., (1993), The structure of the human thyroxine binding globulin (TBG) gene.Biochim. Biophys. Acta 1216:446-454.
  • Hayashi Y., et al.,(1993), Human thyroxine-binding globulin gene: complete sequence and transcriptional regulation.Mol. Endocrinol. 7:1049-1060.
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