FDC-P1 cells were cultured with 0 to 250 ng/mL of hIGF-II. Cell proliferation was assessed after 48 hours by measuring OD490.
The purity of recombinant hIGF-II was determined by SDS-PAGE of 1.5 µg reduced (+) and non-reduced (-) recombinant hIGF-II and staining overnight with Coomassie Blue.
Western blot analysis of extracts from NIH/3T3 cells untreated or treated with hIGF-II for 10 minutes, using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Working concentration of hIGF-II generally ranges from 1-100 ng/ml.
Recombinant human IGF-II is supplied as lyophilized material that is very stable at -20°C. It is recommended to reconstitute with sterile water at a concentration of 0.1 mg/ml which can be further diluted in aqueous solutions as needed. Addition of a carrier protein (0.1% HSA or BSA) is recommended for long term storage.
A greater than 95% purity was determined by SDS-PAGE.
Less than or equal to 1 EU / 1 μg hIGF-II.
The bioactivity of recombinant hIGF-II was determined in a cell proliferation assay using FDC-P1 cells. The ED50 of each lot is between 10-20 ng/ml.
Recombinant human IGF-II was expressed in E. coli and is supplied in a lyophilized form. A greater than 95% purity was determined by SDS-PAGE. Endotoxin levels are less than or equal to 1 EU / 1 μg hIGF-II.
IGF-II is a potent cellular mitogen that is closely related to IGF-I (1). IGF-II is primarily produced by the liver and is frequently overexpressed in tumors (1,2). IGF-II binds to the IGF-IR, activating the AKT, mTOR, ERK, and JNK pathways (1). IGF-II signaling is regulated by several distinct mechanisms. First, IGF binding proteins (IGFBPs) bind to IGF-II and block interactions with the IGF-IR (1-3). Second, the IGF-IIR binds to and acts as a molecular trap for IGF-II (1-3). Lastly, the IGF2 gene is an imprinted gene, and loss of imprinting leads to increased IGF-II levels (1-3). Aberrant levels of IGF-II are associated with Wilms tumor, Beckwith-Wiedmann syndrome, and colorectal cancer (1,2).
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