Product Pathways - Growth Factors/Cytokines
Human Betacellulin (hBTC) #5235
PhosphoSitePlus® protein, site, and accession data: BTC
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Source
Recombinant human Betacellulin (hBTC) Asp31-Tyr111 (Accession #P35070) was produced in E. coli at Cell Signaling Technology.
Molecular Characterization
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.
Purity
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hBTC. All lots are greater than 98% pure.
Bioactivity
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.
Coomassie Gel
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.
Bioactivity
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.
Bioactivity
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.
Endotoxin
Less than 0.01 ng endotoxin/1 μg hBTC.
Formulation
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.
Background
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).
- Dunbar, A.J. and Goddard, C. (2000) Int J Biochem Cell Biol 32, 805-15.
- Oh, Y.S. et al. (2011) PLoS One 6, e23894.
- Yechoor, V. et al. (2009) Endocrinology 150, 4863-73.
- Yamamoto, Y. et al. (2008) Growth Factors 26, 173-9.
Application References
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Companion Products
- 9872 Phosphate Buffered Saline (PBS-1X) pH7.2 (Sterile)
- 7177 PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Kit
For Research Use Only. Not For Use In Diagnostic Procedures.
