The purity of recombinant hBTC was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hBTC and staining overnight with Coomassie Blue.Learn more about how we get our images
The proliferation of MCF 10A cells treated with increasing concentrations of hBTC was assessed. After 24 hr treatment, cells were labeled with BrdU for 4 hr. BrdU incorporation was determined by ELISA and the OD450 was determined.Learn more about how we get our images
The ability of hBTC to induce phosphorylation of p44/42 MAPK (Erk1/2) was assessed. After starvation, MCF 10A cells were treated with increasing concentrations of hBTC for 10 min. Cells were lysed, and Phospho-p44/42 MAPK (Erk1/2) was quantified using PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwhich ELISA Kit #7177. OD450 is shown.Learn more about how we get our images
Recombinant human Betacellulin (hBTC) Asp31-Tyr111 (Accession #P35070) was produced in E. coli at Cell Signaling Technology.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hBTC. All lots are greater than 98% pure.
Recombinant hBTC does not have a Met at the amino terminus and has a calculated MW of 8,984. DTT-reduced and non-reduced protein migrate as 14 kDa polypeptides. The expected amino-terminal DGNST of recombinant hBTC was verified by amino acid sequencing.
The bioactivity of recombinant hBTC was determined in a MCF 10A cell proliferation assay. The ED50 of each lot is between 0.6 and 1.0 ng/ml.
Less than 0.01 ng endotoxin/1 μg hBTC.
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hBTC. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.
Stable in lyophilized state at 4°C for 1 year after receipt. Sterile stock solutions reconstituted with carrier protein are stable at 4°C for 2 months and at -20°C for 6 months. Avoid repeated freeze-thaw cycles. Maintain sterility. Storage at -20°C should be in a manual defrost freezer.
Betacellulin (BTC), a member of the EGF family of ligands, is a widely expressed growth factor that was originally identified in conditioned medium of a mouse pancreatic beta cell line (1). BTC is synthesized as a 178 amino acid transmembrane precursor protein that is proteolytically cleaved into an 80 amino acid mature protein (1). Human BTC shares 74% amino acid sequence identity with mouse BTC (1). BTC can bind to and activate the ErbB family of receptor tyrosine kinases, EGFR/ErbB1, HER2/ErbB2, HER3/ErbB3, and HER4/ErbB4 (1,2). BTC can also bind to and activate EGFR/ErbB1 and HER4/ErbB4 homodimers (1). Ligand-receptor interactions result in the activation of the PI3K and Erk pathways (1). BTC induces the proliferation of pancreatic beta cell lines in vitro (2). Moreover, research studies have shown that BTC induces beta cell regeneration and increased insulin secretion in rodent models of diabetes (2-4).
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