Recombinant Human FGF basic Heat Stable GMP Protein, CF

Catalog #: BT-FGFBHS-GMP Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
BT-FGFBHS-GMP-025
BT-FGFBHS-GMP-01M
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Recombinant Human FGF basic Heat Stable GMP Protein Bioactivity.
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Recombinant Human FGF basic Heat Stable GMP Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. 17192 Da ± 20 Da
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
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 0.0500-0.600 ng/mL.  
The specific activity of Recombinant Human FGF basic Heat Stable GMP is >1.6 x 106 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 #
N-terminal Sequence
Analysis
Ala135-Ala-Gly-Ser-Ile-Thr-Thr-Leu-Pro-Ala & Ala136-Gly-Ser-Ile-Thr-Thr-Leu-Pro-Ala-Leu
Predicted Molecular Mass
17 kDa
SDS-PAGE
16-19 kDa, under reducing conditions.

Product Datasheets

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BT-FGFBHS-GMP

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.

BT-FGFBHS-GMP

Formulation Lyophilized from a 0.2 μm filtered solution in HEPES and Sodium Sulfate with Trehalose.
Reconstitution Reconstitute the 25 μg size at 250 μg/mL in sterile water. Reconstitute all other sizes at 500 μg/mL in sterile water.
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.
  • 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.

Scientific Data

Bioactivity View Larger

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.

Bioactivity View Larger

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).

Cell Culture View Larger

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.

Cell Culture View Larger

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.

SDS-PAGE View Larger

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.

Reconstitution Calculator

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. 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. 

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 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

Manufacturing Specifications

GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

R&D Systems' quality focus includes:

  • Designed, manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
  • Documented and controlled manufacturing process
  • Control of documentation and process changes by QA
  • Personnel training programs
  • Raw material inspection and vendor qualification/monitoring program
  • Validated equipment, processes and test methods
  • Equipment calibration and maintenance schedules using a Regulatory Asset Manager
  • Facility/Utilities maintenance, contamination controls, safety and pest control programs
  • Material review process for variances
  • Robust product stability program following relevant ICH guidelines

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their application. Each product is provided with a lot-specific Certificate of Analysis that contains the product's specifications and test results. Quality control testing may include, but is not limited to:

  • N-terminal amino acid analysis
  • SDS-PAGE purity analysis
  • Molecular weight analysis via mass spectrometry
  • Endotoxin assessment per USP <85> and Ph. Eur. 2.6.14 guidelines
  • Bioassay analysis
  • Microbial testing per USP <71> and Ph. Eur. 2.6.1 guidelines
  • Host cell protein assessment
  • Host cell DNA assessment
  • Mycoplasma assessment

Additional testing and documentation requested by the customer can be arranged at an additional cost. 

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis and St. Paul, Minnesota USA.

R&D Systems sells GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.


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.

    Please read our complete Animal-Free Statement

    Product Specific Notices

    Produced under license from Enantis s.r.o. Full terms and conditions of sale can be found online in the Protein Sciences Segment T&Cs at: Terms & Conditions.

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