Application Note
INS (Insulin) is a peptide hormone produced by beta cells of the pancreatic islets, which decreases blood glucose concentration and increases cell permeability to monosaccharides, amino acids and fatty acids. It has been reported that insulin triggers phosphorylation of a number of substrates by binding to its receptors, which was important for cell proliferation, cell cycle progression, cell division and differentiation. To detect the effect of Insulin on cell proliferation, MCF-7 cells were seeded into triplicate wells of 96-well plates at a density of 2000 cells/well and allowed to attach overnight, then the medium was replaced with serum-free standard DMEM prior to the addition of various concentrations of INS. After incubated for 48h, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10 μl of CCK-8 solution was added to each well of the plate, then the absorbance at 450nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37ºC. The dose-effect curve showed INS significantly promoted cell proliferation of MCF-7 cells. The ED₅₀ for this effect is typically 11. 71-57. 11 ng/ml.
Observed MW
10 kDa.
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Phe25~Ser54(B)+GGGGS+Gly90~Asn110(A) (NP_001172012.1)
Expression System
E. coli
Purity
> 97%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Synonyms
insulin II , AA986540 , Ins-2 , InsII , Mody , Mody4
Background
This gene encodes insulin, a peptide hormone that plays a vital role in the regulation of carbohydrate and lipid metabolism. The encoded precursor protein undergoes proteolytic cleavage to produce a disulfide-linked heterodimeric functional protein that is stored in secretory granules. An increase in blood glucose levels, among others, induces the release of insulin from the secretory granules. Mice deficient in the functional hormone encoded by this gene develop diabetes mellitus. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Aug 2015]
Database
Research Area