Recombinant Human SCF GMP Protein, CF GMP
R&D Systems, part of Bio-Techne | Catalog # 255B-GMP
Animal-Free
Key Product Details
Product Specifications
Source
E. coli-derived human SCF/c-kit Ligand protein
Glu26-Ala189, with an N-terminal Met.
Produced using non-animal reagents in an animal-free laboratory.
Manufactured and tested under cGMP guidelines.
Glu26-Ala189, with an N-terminal Met.
Produced using non-animal reagents in an animal-free laboratory.
Manufactured and tested under cGMP guidelines.
Purity
>97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. The result from mass spectrometry confirms the major peak corresponds to SCF/c-kit ligand. See the CofA for additional information.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Met-Glu26-Gly-Ile-(Cys)-Arg-Asn-Arg-Val-Thr
Predicted Molecular Mass
19 kDa
SDS-PAGE
17-18 kDa, reducing conditions
Activity
Measured in a cell proliferation assay using TF-1 human erythroleukemic cells. Kitamura, T. et al. (1989) J. Cell Physiol. 140:323.
The ED50 for this effect is 1‑5 ng/mL.The specific activity of recombinant human SCF is >1.0 x 106 units/mg, which is calibrated against the human SCF reference standard (NIBSC code: 91/682).
The ED50 for this effect is 1‑5 ng/mL.The specific activity of recombinant human SCF is >1.0 x 106 units/mg, which is calibrated against the human SCF reference standard (NIBSC code: 91/682).
Host Cell Protein
<0.1 ng per µg of protein when tested by ELISA.
Mycoplasma
Negative for Mycoplasma.
Host Cell DNA
<0.010 ng per µg of protein when tested by PCR.
Scientific Data Images for Recombinant Human SCF GMP Protein, CF
Recombinant Human SCF GMP Protein Bioactivity
GMP-grade Recombinant Human SCF (Catalog # 255B-GMP) stimulates cell proliferation of the TF‑1 human erythroleukemic cell line. The ED50 for this effect is 1-5 ng/mL. Four independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP SCF.
Equivalent Bioactivity of GMP and Animal-Free grades of Recombinant Human SCF
Equivalent bioactivity of GMP (Catalog # 255B-GMP) and Animal-Free (AFL255) grades of Recombinant Human SCF as measured in cell proliferation assay (orange and green respectively).
Recombinant Human SCF GMP Protein SDS-PAGE
2 μg/lane of GMP-grade Recombinant Human SCF (Catalog # 255B-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing R band at 18 kDa and NR band at 16 kDa, respectively.Formulation, Preparation and Storage
255B-GMP
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS. |
| 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.
|
Background: SCF/c-kit Ligand
Stem cell factor (SCF), also known as c-kit ligand (KL), mast cell growth factor (MGF), and steel factor (SLF), is a widely expressed 28‑40 kDa type I transmembrane glycoprotein (1). It promotes the survival, differentiation, and mobilization of multiple cell types including myeloid, erythroid, megakaryocytic, lymphoid, germ cell, and melanocyte progenitors (1‑7). SCF is a primary growth and activation factor for mast cells and eosinophils (8). Mature human SCF consists of a 189 amino acid (aa) extracellular domain (ECD), a 23 aa transmembrane segment, and a 36 aa cytoplasmic tail (9). The ECD shows both N‑linked and O-linked glycosylation (10). Proteolytic cleavage at two alternate sites in the extracellular juxtamembrane region releases a 25 kDa soluble molecule which is comparable to the only form produced by Steel-dickie mutant mice (11, 12). An alternately spliced isoform of human SCF lacks 28 aa that encompasses the primary proteolytic recognition site (13). Within the ECD of the long isoform (corresponding to this recombinant protein), human SCF shares 79%‑87% aa sequence identity with canine, feline, mouse, and rat SCF. Rat SCF is active on mouse and human cells, but human SCF is only weakly active on mouse cells (9). Noncovalent dimers of transmembrane or soluble SCF interact with the receptor tyrosine kinase SCF R/c‑kit to trigger receptor dimerization and signaling (14). SCF assists in the recovery of cardiac function following myocardial infarction by increasing the number of cardiomyocytes and vascular channels (15).
References
- Ashman, L.K. (1999) Int. J. Biochem. Cell Biol. 31:1037.
- Sette, C. et al. (2000) Int. J. Dev. Biol. 44:599.
- Yoshida, H. et al. (2001) J. Invest. Dermatol. Symp. Proc. 6:1.
- Erlandsson, A. et al. (2004) Exp. Cell Res. 301:201.
- Kapur, R. et al. (2002) Blood 100:1287.
- Wang, C.-H. et al. (2007) Arterioscler. Thromb. Vasc. Biol. 27:540.
- Bashamboo, A. et al. (2006) J. Cell Sci. 119:3039.
- Reber, L. et al. (2006) Eur. J. Pharmacol. 533:327.
- Martin, F.H. et al. (1990) Cell 63:203.
- Arakawa, T. et al. (1991) J. Biol. Chem. 266:18942.
- Majumdar, M.K. et al. (1994) J. Biol. Chem. 269:1237.
- Brannan, C.I. et al. (1991) Proc. Natl. Acad. Sci. 88:4671.
- Anderson, D.M. et al. (1991) Cell Growth Differ. 2:373.
- Lemmon, M.A. et al. (1997) J. Biol. Chem. 272:6311.
- Kanellakis, P. et al. (2006) Cardiovasc. Res. 70:117.
Long Name
Stem Cell Factor
Alternate Names
c-kit Ligand, DCUA, DFNA69, FPH2, FPHH, KITLG, KL-1, MGF, SHEP7, SLF
Gene Symbol
KITLG
UniProt
Additional SCF/c-kit Ligand Products
Product Documents for Recombinant Human SCF GMP Protein, CF
Manufacturing Specifications
GMP ProteinsR&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:
- Manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
- Documented processes and QA control of documentation and process changes
- Personnel training programs
- Raw material testing and vendor qualification/monitoring
- Fully validated equipment, processes and test methods
- Equipment calibration schedules using a computerized calibration program
- Facility maintenance, safety programs and pest control
- Material review process for variances
- Monitoring of stability over product shelf-life
- N-terminal amino acid analysis, SDS-PAGE analysis, mass spectrometry, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
- Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial testing according to USP
- Host Cell Protein testing performed by ELISA
- Mycoplasma testing by ribosomal RNA hybridization assay
Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis, 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.
- 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.
- Low Endotoxin Level.
- No impairment of biological activity.
- High quality product obtained under stringent conditions.
Product Specific Notices for Recombinant Human SCF GMP Protein, CF
END USER TERMS OF USE OF PRODUCT
The following terms are offered to you upon your acceptance of these End User Terms of Use of Product. By using this product, you indicate your acknowledgment and agreement to these End User Terms of Use of Product. If you do not agree to be bound by and comply with all of the provisions of these End User Terms of Use of Product, you should contact your supplier of the product and make arrangements to return the product.
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The End User is aware that R&D Systems, Inc. sells GMP products for preclinical or clinical ex vivo use and not for in vivo use. The End User further agrees, as a condition of the sale of R&D Systems' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be directly or indirectly administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.
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You agree to review these End User Terms of Use of Product to ensure any subsequent use by you of R&D Systems' GMP Products following changes to these End User Terms of Use of Product constitutes your acceptance of all such changes.
TERMS AND CONDITIONS
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For preclinical, or clinical ex vivo use
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