Product Description
This Human Resistin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Resistin protein (Cat: 13055-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human RETN (NP_065148.1) (Ser17-Pro108) was expressed with the Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human RETN consists 352 amino acids and predicts a molecular mass of 38.2 kDa.
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 ADSF Overexpression Lysate;Human FIZZ3 Overexpression Lysate;Human RETN1 Overexpression Lysate;Human RSTN Overexpression Lysate;Human XCP1 Overexpression Lysate
Resistin is an adipocytokine, which has been studied for its role in insulin resistance and recently in inflammation. The RETN and CAP1 polymorphisms and gene expression may be potential biomarkers for breast cancer risk. Resistin (RETN), recently found to be relevant to inflammation and inflammatory disorders.
References
Verma S, et al. (2003) Resistin promotes endothelial cell activation: further evidence of adipokine-endothelial interaction. Circulation. 108(6):736-40. Reilly MP, et al. (2005) Resistin is an inflammatory marker of atherosclerosis in humans. Circulation. 111(7):932-9. Steppan CM, et al. (2002) Resistin and obesity-associated insulin resistance. Trends Endocrinol Metab. 13(1):18-23.