Recombinant Human FGF basic/FGF2/bFGF GMP Protein, CF

GMP-grade Recombinant Human FGF basic/FGF2 Protein (Catalog # BT-FGFB-GMP) as measured in a cell proliferation assay using NR6R-3T3 mouse fibroblast cell line. The ED50 for this effect is 0.100-1.00 ng/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP FGF basic/FGF2.

Equivalent bioactivity of GMP (Catalog # BT-FGFB-GMP) and Animal-Free (BT-FGFB-AFL) grades of Recombinant Human FGF basic/FGF2 as measure in a cell proliferation assay using NR6R-3T3 mouse fibroblast cell line (orange and green, respectively).

2 μg/lane of Recombinant Human FGF basic/FGF2/bFGF GMP Protein (Catalog # BT-FGFB-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 18 kDa, under reducing conditions.

iPSCs were cultured for five passages in a commercially available media without FGF2 + 100µg/mL of Recombinant Human GMP FGF basic (Catalog # BT-FGFB-GMP). Cells were analyzed for expression of stemness markers APC-conjugated OCT-3/4 (Catalog # ICA1759A) and PE-conjugated Nanog (Catalog # IC1997P) via flow cytometry. Most cells expressed high levels of OCT-3/4 and Nanog (≥95%).

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

Recombinant Human FGF basic/FGF2/bFGF 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 ED₅₀ for this effect is 0.100-1.00 ng/mL.
  The specific activity of Recombinant Human FGF basic/FGF2 GMP is >8.0 x 10⁵ 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
  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: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The primary molecular weight by mass spectrometry is 17112 Da ± 10 Da
• Endotoxin Note: <0.01 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: 18 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 °C as supplied. Refer to lot specific COA for the Use by Date.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in HEPES and Sodium Sulfate with Trehalose.
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The primary molecular weight by mass spectrometry is 17112 Da ± 10 Da
• Reconstitution Instructions: Reconstitute 25 μg size at 250 μg/mL and all other sizes at 500 μg/mL in sterile water.

Notes

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Alternate Names for Recombinant Human FGF basic/FGF2/bFGF 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 FGF2 and HBGF-2) is an 18-34 kDa, heparin-binding member of the FGF superfamily of molecules (1-3). Superfamily members are characterized by the presence of a centrally placed beta -trefoil structure. FGF acidic (FGF-1) and FGF basic (FGF2) were the first two identified FGFs, and the designations acidic and basic refer to their relative isoelectric points. Human FGF basic is 288 amino acids (aa) in length. There are multiple start sites, four of which utilize atypical CUG codons, and one that initiates at an AUG start site (4 - 6). The four CUG start sites generate high molecular weight (HMW) FGF basic. There is a 34 kDa, 288 aa form, a 24 kDa, 210 aa form, a 22.5 kDa, 201 aa form, and a 22 kDa, 196 aa form. All are retained intracellularly, undergo extensive methylation, and possess one or more nuclear localization signals (NLS) (7-9). The AUG initiating form is 18 kDa and 155 aa in length. There is no signal sequence (ss). It is, however, secreted directly through the plasma membrane via a mechanism that appears to be dependent upon tertiary structure (10). In place of a ss, there is purportedly a 9 aa N-terminal prosegment that precedes a 146 aa mature segment (11). Early isolations of 18 kDa bovine FGF basic yielded 146 aa molecules, an effect attributed to the presence of acid proteases (12). The molecule contains a heparin-binding site (aa residues 128-144), and undergoes phosphorylation at Ser117 (13). There is also an ill-defined C-terminal NLS that may be more “functional” (or 3-dimensional) than structural (7). Human 146 aa FGF basic is 97% aa identical to mouse FGF basic (14).

1. Sorenson, V. et al. (2006) BioEssays 28:504.
2. Kardami, E. et al. (2004) Cardiovasc. Res. 63:458.
3. Nugent, M.A. and R.V. Lozzo (2000) Int. J. Biochem. Cell Biol. 32:115.
4. Abraham, J.A. et al. (1986) EMBO J. 5:2523.
5. Prats, H. et al. (1989) Proc. Natl. Acad. Sci. USA 86:1836.
6. Arnaud, E. et al. (1999) Mol. Cell. Biol. 19:505.
7. Foletti, A. et al. (2003) Cell. Mol. Life Sci. 60:2254.
8. Arese, M. et al. (1999) Mol. Biol. Cell 10:1429.
9. Pintucci, G. et al. (1996) Mol. Biol. Cell 7:1249.
10. Nickel, W. (2005) Traffic 6:607.
11. SwissProt # P09038.
12. Klagsbrun, M. et al. (1987) Proc. Natl. Acad. Sci. USA 84:1839.
13. Bailly, K. et al. (2000) FASEB J. 14:333.
14. Hebert, J.M. et al. (1990) Dev. Biol. 138:454.

FAQs for FGF basic/FGF2/bFGF (BT-FGFB-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)