Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein

Name: Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein
Brand: R&D Systems
SKU: AFL3718

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AFL3718

Also available, GMP FGF basic/FGF2/bFGF Protein, Catalog # 3718-GMP! Animal-free and manufactured under GMP guidelines for therapeutic manufacturing applications.

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Source:	E. coli
Accession #:	NP_001997.5
Applications:	Bioactivity

View Full Technical Details for Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein

Product Specifications for Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein

Source	E. coli-derived human FGF basic/FGF2/bFGF protein Pro143-Ser288 Produced using non-animal reagents in an animal-free laboratory.
Purity	>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin level	<0.01 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Pro143 & Ala144
Predicted Molecular Mass	16 kDa
Activity	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 0.1-0.6 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).

Scientific Data Examples for Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein

	Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein Bioactivity Animal-Free^TM Recombinant Human FGF basic/FGF2/bFGF (Catalog # AFL3718) stimulates proliferation of the NR6R-3T3 mouse fibroblast cell line. The ED₅₀ for this effect is 0.1-0.6 ng/mL.
	Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein SDS-PAGE 2 μg/lane of Recombinant Human FGF basic Animal-Free Protein (Catalog #AFL3718) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 17 kDa and 18 kDa, respectively.
	Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein SDS-PAGE. 1 μg/lane of Animal-Free^TM Recombinant Human FGF basic/FGF2/bFGF Protein (Catalog # AFL3718) was resolved with SDS-PAGE under reducing (R) conditions and non-reducing (NR) conditions and visualized by silver staining, showing a single band at 17 kDa.

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AFL3718

Formulation	Lyophilized from a 0.2 μm filtered solution in Tris-HCl and NaCl.
Reconstitution	Reconstitute at 100 μg/mL in PBS.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: FGF basic/FGF2/bFGF

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).

Item	Value
References	Sorenson, V. et al. (2006) BioEssays 28:504. Kardami, E. et al. (2004) Cardiovasc. Res. 63:458. Nugent, M.A. and R.V. Lozzo (2000) Int. J. Biochem. Cell Biol. 32:115. Abraham, J.A. et al. (1986) EMBO J. 5:2523. Prats, H. et al. (1989) Proc. Natl. Acad. Sci. USA 86:1836. Arnaud, E. et al. (1999) Mol. Cell. Biol. 19:505. Foletti, A. et al. (2003) Cell. Mol. Life Sci. 60:2254. Arese, M. et al. (1999) Mol. Biol. Cell 10:1429. Pintucci, G. et al. (1996) Mol. Biol. Cell 7:1249. Nickel, W. (2005) Traffic 6:607. SwissProt # P09038. Klagsbrun, M. et al. (1987) Proc. Natl. Acad. Sci. USA 84:1839. Bailly, K. et al. (2000) FASEB J. 14:333. Hebert, J.M. et al. (1990) Dev. Biol. 138:454.
Long Name	Fibroblast Growth Factor basic
Entrez Gene IDs	2247 (Human); 14173 (Mouse); 281161 (Bovine); 403857 (Canine); 100033955 (Equine)
Alternate Names	Basic fibroblast growth factor, FGF basic, FGF-2, FGF2, FGFBprostatropin, HBGF-2, Prostatropin, bFGF, basic fibroblast growth factor bFGF, fibroblast growth factor 2 (basic), heparin-binding growth factor 2

Manufacturing Specifications

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.

Production

All molecular biology procedures use animal-free media and dedicated labware.
Dedicated fermentors are utilized in committed animal-free areas.

Purification

Protein purification columns are animal-free.
Bulk proteins are filtered using animal-free filters.
Purified proteins are stored in animal-free containers in a dedicated cold storage room.

Quality Assurance

Low Endotoxin Level.
No impairment of biological activity.
High quality product obtained under stringent conditions.
For ex vivo research or bioproduction, additional documentation can be provided.

Please read our complete Animal-Free Statement

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Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein

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Product Specifications for Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein

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Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: FGF basic/FGF2/bFGF

Manufacturing Specifications

Citations for Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein

Product Documents for Recombinant Human FGF basic/FGF2/bFGF, Animal-Free Protein

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