Recombinant Human FGF basic Heat Stable GMP Protein, CF

The bioactivity of Recombinant Human FGF basic Heat Stable GMP Protein (Catalog # BT-FGFBHS-GMP) was measured in a cell proliferation assay using NR6R-3T3 mouse fibroblast cell line. The ED50 for this effect is 0.0500-0.600 ng/mL. Three independent lots were tested for bioactivity and plotted on the same graph to show lot-to-lot consistency of GMP FGF basic/FGF2 Heat Stabile protein.

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

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/cm2in a 6-well dish coated with Cultrex Ultimatrix (BME001). 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.

2 μg/lane of Recombinant Human FGF basic Heat Stable GMP Protein (Catalog # BT-FGFBHS-GMP) 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.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human FGF basic Heat Stable GMP Protein, CF Summary

• Details of Functionality: Measured in a cell proliferation assay using NR6R-3T3 mouse fibroblast cells. Raines, E.W. et al. (1985) Methods Enzymol. 109:749. The ED 50 for this effect is 0.0500-0.600 ng/mL. The specific activity of Recombinant Human FGF basic Heat Stable GMP is >1.6 x 10 6 IU/mg, which is calibrated against the human FGF basic/FGF2 WHO International Standard (NIBSC code: 90/712).
• Source: E. coli-derived human FGF basic/FGF2/bFGF protein Ala135-Ser288 (with modifications) Produced using non-animal reagents in an animal-free laboratory. Manufactured and tested under cGMP guidelines.
• Accession #: NP_001997.5
• N-terminal Sequence: Ala135-Ala-Gly-Ser-Ile-Thr-Thr-Leu-Pro-Ala & Ala136-Gly-Ser-Ile-Thr-Thr-Leu-Pro-Ala-Leu
• Protein/Peptide Type: GMP Recombinant Proteins
• Purity: >95%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. 17192 Da ± 20 Da
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 17 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 16-19 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  A minimum of 12 months when stored at ≤ -20 degreesC as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 3 months, ≤ -20 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in HEPES and Sodium Sulfate with Trehalose.
• Purity: >95%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. 17192 Da ± 20 Da
• Reconstitution Instructions: Reconstitute the 25 μg size at 250 μg/mL in sterile water. Reconstitute all other sizes at 500 μg/mL in sterile water.

Notes

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Alternate Names for Recombinant Human FGF basic Heat Stable GMP Protein, CF

• basic fibroblast growth factor bFGF
• Basic fibroblast growth factor
• bFGF
• FGF basic
• FGF2
• FGF-2
• FGFBprostatropin
• fibroblast growth factor 2 (basic)
• HBGF-2
• heparin-binding growth factor 2
• Prostatropin

Background

FGF basic (also known as FGF-2 and HBGF-2) is an 18-34 kDa, heparin-binding member of the FGF superfamily of molecules. As a growth factor, FGF basic is expressed in a variety of cell types and involved in the regulation of cell proliferation, differentiation, migration and survival. FGF basic promotes self-renewal and upregulates pluripotency markers such as Oct4, Sox2 and Nanog, making it well-known for maintenance of pluripotency in embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). These functions make FGF basic an important cell culture supplement in stem cell applications for regenerative medicine and clinical manufacturing protocols. Our Heat Stable FGF basic has been engineered for thermostability and maintains its activity at 37℃ when compared to wild-type. Heat-stable FGF basic can enhance the flexibility and efficiency of cell culture by reducing the need for frequent media changes.

FAQs for FGF basic/FGF2/bFGF (BT-FGFBHS-GMP). (Showing 1 - 1 of 1 FAQ).

1. I am looking for an ovine antibody for bFGF and PDGF-AB and ELISA Kits for both. It is for a research work on meniscus healing in sheep. Do you have anything suitable? How is the homology with sheep? Is it ever tested and/or published in papers?
   • Unfortunately, most of the products you are interested in have not yet been tested in sheep, but this should not be a problem, due to sequence homology, and our Innovators Reward Program. In terms of an antibody for bFGF, also called FGF-2, we have several antibodies that are validated to detect the bovine protein, which is almost identical to that from sheep (99% homology). We only have FGF2 ELISA kits that have been tested with human or mouse. Mouse FGF-2 has 94% homology with sheep; human has 98% homology for most of its sequence, but also has an additional stretch of amino acids that is not present in the sheep protein. I therefore think that the mouse kit would be most suitable, just in case the human kit targets the region of the human protein not present in mouse. We sell antibodies to PDGF-A or PDGF-B rather than PDGF-AB. We don't have any PDGF-A antibodies that have been tested in sheep but there are several options you may find of interest. There isn't a sheep PDGFA sequence in UniProt, so I can't comment on how similar it is to the PDGFA from other species. Nevertheless, our NBP1-52533 PDGFB antibody has been shown to work in sheep. We sell two PDGF-AB ELISA kits , but these are for human and rat. I'm not sure how well these would work with sheep. Note that if you test a product with a species (or application) that it has not yet been validated in, you are eligible for our Innovator's Reward: Novus would provide you a 50% refund on the purchased product as well as a 50% discount on a future product of equal or lesser value. You would email innovators@novusbio.com to apply for your award, and we

Bioinformatics

• Uniprot:
  • NP_001997.5(Human)