Recombinant Zebrafish FGF-2 (bFGF), Animal-Free Protein

Zebrafish FGF2 activity is determined using the Promega serum response element luciferase reporter assay (*) in transfected HEK293T cells.Cells are treated in triplicate with a serial dilution of FGF2 for 6 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. EC50 = 0.42 ng/ml (24.7 pM).*Promega pGL4.33[luc2P/SRE/Hygro] #E1340

FGF2 migrates as major band at 17 kDa in non-reducing (-beta ME) conditions and upon reduction (+ beta ME). The higher molecular mass band at 35 kDa is a dimer that we always see in our highly purified zebrafish FGF2 protein, the presence of this does not affect biological activity. Purified recombinant protein (7 µg) was resolved using 15% w/v SDS-PAGE in reduced (+ beta -mercaptoethanol, R) and non-reduced conditions (NR) and stained with Coomassie Brilliant Blue R250.

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

Recombinant Zebrafish FGF-2 (bFGF), Animal-Free Protein Summary

• Details of Functionality: No significant difference between EC₅₀ of reference and test lots
• Source: E. coli-derived zebrafish FGF basic/FGF2/bFGF protein
• Accession #: AAI62367.1
• Protein/Peptide Type: Animal-Free Recombinant Proteins
• 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: Monomeric zebrafish FGF-2 (bFGF) protein only

Packaging, Storage & Formulations

• Storage: Store lyophilized protein between -20 and -80 °C until the date of expiry. Avoid freeze-thaw cycles.
• Buffer: Lyophilized from Tris/NaCl/CyS/mannitol
• Reconstitution Instructions: Resuspend in water at >100 µg/ml, prepare single use aliquots, add carrier protein if desired.

Notes

The above product was manufactured, tested and released by R&D System's contract manufacturer, Qkine Ltd, at 1 Murdoch House, Cambridge, UK, CB5 8HW. The product is for research use only and not for the diagnostic or theraputic use.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Zebrafish FGF-2 (bFGF), Animal-Free Protein

• 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 (1-3). Superfamily members are characterized by the presence of a centrally placed beta -trefoil structure. FGF acidic (FGF-1) and FGF basic (FGF-2) 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 (Qk002). (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:
  • AAI62367.1()