Recombinant Human KGF/FGF-7 GMP Protein, CF

Catalog #: BT-KGF-GMP Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
BT-KGF-GMP-050
BT-KGF-GMP-01M

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Recombinant Human KGF/FGF‑7 GMP Protein Bioactivity.

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human KGF/FGF-7 GMP Protein, CF Summary

KGF/FGF-7 Manufactured in Bio-Techne's new GMP facility
Lot-to-lot consistency
Stringent guidelines for patient safety
Scalability necessary to support successful therapeutics
Learn more about manufacturing in our new GMP facility
Test it in your process! Request a sample of GMP KGF

Product Specifications

Purity

>97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 18927 Da ± 61 Da

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured in a cell proliferation assay using Ba/F3 mouse pro B cells transfected with human FGF RIIb. The ED₅₀ for this effect is 6.00-60.0 ng/mL.

The specific activity of recombinant human KGF/FGF-7 is >8.16 x 10⁵ units/mg, which is calibrated against the human KGF/FGF-7 WHO Standard (NIBSC code: 03/150).

Source

E. coli-derived human KGF/FGF-7 protein
Cys32 - Thr194, with an N-terminal Met
Produced using non-animal reagents in an animal-free laboratory.

Manufactured and tested under cGMP guidelines.

Accession #

P21781.1

N-terminal Sequence
Analysis

(Cys)₃₂-Asn-Asp-Met-Thr-Pro-Glu-Gln-Met-Ala

Asn₃₃-Asp-Met-Thr-Pro-Glu-Gln-Met-Ala-Thr

Predicted Molecular Mass

19 kDa

SDS-PAGE

20 kDa, under reducing conditions.

Product Datasheets

BT-KGF-GMP

Product Datasheet

COA

SDS

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-KGF-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in MOPS, Na₂SO₄ and EDTA with Trehalose.
Reconstitution	Reconstitute 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

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The bioactivity of Recombinant Human KGF/FGF‑7 GMP Protein (Catalog # BT-KGF-GMP) was measured in a cell proliferationassay using Ba/F3 mouse pro B cells transfected with human FGF RIIb. The ED₅₀ for this effect is 6.00 ‑ 60.0 ng/mL. Threeindependent lots were tested for bioactivity and plotted on the same graph to show lot-to-lotconsistency of GMP KGF/FGF‑7 protein.

Bioactivity

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Equivalent bioactivity of GMP(Catalog # BT-KGF-GMP) andAnimal-Free (BT-KGF-AFL) grades of RecombinantHuman KGF/FGF-7 measured in a cell proliferationassay using Ba/F3 mouse pro Bcells transfected with human FGFRIIb (orange and green, respectively).

SDS-PAGE

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2 μg/lane of Recombinant Human KGF/FGF‑7 GMP Protein (Catalog # BT-KGF-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 20 kDa, under reducing conditions.

Data

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Characterization of the pancreatic progenitor cells differentiated from iPSCs using different grades of Recombinant Human KGF Protein (Catalog # BT-KGF, BT-KGF-AFL, and BT-KGF-GMP). The iPSC cells (1210a cell line) were cultured in a 24-well plate and differentiated into pancreatic progenitor cells using different lots of KGF and other differentiation factors. Expression of pancreatic progenitor markers was analyzed via flow cytometry with anti-PDX-1 (Catalog # MAB2419) and anti-NKX6.1 antibodies. The PDX-1+/NKX6.1+ double positive cell population was quantified as the pancreatic progenitor cells. The data is represented as the average of three technical replicates.

Data

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Characterization of the endocrine cells and beta islets differentiated from iPSCs using Recombinant Human KGF/FGF-7 GMP Protein (Catalog # BT-KGF-GMP). A) Brightfield images of the endocrine cells and beta islets. GMP KGF was used for further differentiation of the pancreatic progenitor cells first into endocrine cells and then beta islets. The endocrine cells and beta islets were imaged with an EVOS Cell Imaging System. B) The expression of beta islet markers was analyzed via flow cytometry with anti-NKX6.1 and anti-C-peptide antibodies. The C-peptide+/NKX6.1+ double positive cell population was quantified as the target beta cells. The data is represented as the average of three technical replicates.

Data

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Immunofluorescent images of the beta islets generated using Recombinant Human KGF/FGF-7 GMP Protein (Catalog # BT-KGF-GMP). Beta islets differentiated from iPSCs were fixed, frozen and sectioned. Sections were stained with the anti-C-peptide antibody (Catalog # MAB14171) and the anti-somatostatin antibody (Catalog # MAB2358), followed by the secondary antibody staining (anti-mouse IgG NL493-conjugated antibody: Catalog # NL009; anti-rat IgG NL557-conjugated antibody: Catalog # NL013). A Leica THUNDER Imaging System was utilized for the imaging of C-peptide and somatostatin expression in the beta islets. GMP KGF successfully differentiates the iPSCs into functional beta islets capable of expressing these characteristic markers.

Data

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The insulin secretion function of the iPSC-derived beta islets was tested in a glucose-stimulated insulin secretion assay using the Ella™ Automated ELISA platform, and the cell culture supernatant was analyzed with the Ella™ Automated ELISA platform and Simple Plex human insulin cartridge (Catalog # SPCKC-CS-008554). After the incubation of beta islets with 20 mM glucose for 1 hour at 37 °C, the beta islets generated using Recombinant Human KGF/FGF-7 GMP Protein (Catalog # BT-KGF-GMP) were shown to secrete insulin. The data is represented as the average of three technical replicates.

Reconstitution Calculator

Background: KGF/FGF-7

Keratinocyte Growth Factor (KGF), or Fibroblast Growth Factor-7 (FGF-7), is an important component in many cell culture protocols. It is widely used to promote the growth and differentiation of epithelial cells, including skin keratinocytes, and lung, corneal, and intestinal epithelia. It is a part of tissue engineering and organoid culture protocols, where it can enhance the generation of complex tissues such as liver, gastrointestinal tract, lung, and mammary gland. Furthermore, KGF/FGF-7 has been shown to stimulate the proliferation of pancreatic endocrine progenitor cells and promote their differentiation into mature insulin-secreting beta cells. In these capacities KGF/FGF-7 has significant clinical relevance in studies to better understand gastrointestinal disorders, wound healing, respiratory disease, and diabetes.

Long Name

Keratinocyte Growth Factor

Entrez Gene IDs

2252 (Human); 403915 (Canine)

Alternate Names

FGF7; FGF-7; fibroblast growth factor 7HBGF-7; HBGF7; HBGF-7; Heparin-binding growth factor 7; keratinocyte growth factor; KGF; KGFfibroblast growth factor 7 (keratinocyte growth factor)

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