Recombinant Human FGF basic Heat Stable Protein, CF

Recombinant Human FGF basic Heat Stable (Catalog # BT-FGFBHS ) has a molecular weight (MW) of 20.2 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer.

Equivalent bioactivity of GMP (BT-FGFBHS-GMP), Animal-Free (BT-FGFBHS-AFL) and RUO (Catalog # BT-FGFBHS) grades of Recombinant Human FGF basic Heat Stable proteins as measured in a cell proliferation assay using NR6R-3T3 mouse fibroblast cells (orange, green, red, respectively).

Recombinant Human FGF basic Heat Stable Protein induces NR6R 3T3 mouse fibroblast cell proliferation. (A) Recombinant Human FGF basic Heat Stable Protein (Catalog # BT-FGFBHS) or (B) wild type (WT) Recombinant Human FGF basic/FGF2/bFGF Proteins were untreated or incubated at 37°C for 3 days in media. Human FGF basic Heat Stable (HS) retained similar activity to the untreated HS protein indicating that the HS protein has increased thermal stability. In contrast, WT Recombinant Human FGF basic/FGF2/bFGF Protein significantly lost activity suggesting less thermal stability.

2 μg/lane of Recombinant Human FGF basic Heat Stable Protein (Catalog # BT-FGFBHS) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 16-19 kDa, under reducing conditions.

Human iPSCs were cultured for four passages in a commercial iPSC Expansion media containing 40 ng/mL of either rhFGFb Heat Stable (BT-FGFBHS) or wild type rhFGFb, Animal-Free (BT-FGFB-AFL). Media exchanges were performed following two feeding schedules. A). a daily feed schedule or B). a reduced feed schedule, where cells were double fed twice per week. Cells were plated at 5000 cells/cm² in a 6-well dish coated with Cultrex Ultimatrix (BME001-05). For each schedule, all conditions were passaged once either group reached ~60-70% confluency and counts were performed using a Cellaca™ High-Throughput Cell Counter. Count of each passage is shown comparing WT vs HS FGFb in both the A) daily feed or B) reduced feed schedule.

Human iPSCs were cultured for four passages in a commercial iPSC Expansion media containing 100 ng/mL of either rhFGFb Heat Stable (BT-FGFBHS) or wild type rhFGFb, Animal-Free (BT-FGFB-AFL). Media exchanges were performed following a reduced feed schedule, where cells were double fed twice per week. Cells were plated at 5000 cells/cm² in a 6-well dish coated with Cultrex Ultimatrix (BME001-05). Cells were passaged at ~60-70% confluency and counts were performed using a Cellaca™ High-Throughput Cell Counter. Count of each passage is shown comparing WT vs HS FGFb.