Product Specifications for Recombinant Human BMP-4 GMP Protein, CF
Source
Mouse myeloma cell line, NS0-derived human BMP-4 protein Ser293-Arg408 Manufactured and tested under current Good Manufacturing Practice (GMP) guidelines.
Purity
>95%, by SDS-PAGE with silver staining, under reducing conditions.
Endotoxin level
<0.01 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Ser293-Pro-Lys-His-His-Ser-Gln-Arg-Ala-Arg
Predicted Molecular Mass
13 kDa (monomer)
SDS-PAGE
13 kDa (monomer)
Activity
Measured by its ability to induce alkaline phosphatase production by ATDC5 mouse chondrogenic cells. Binnerts, M.E. et al. (2004) Biochem. Biophys. Res. Commun. 315(2):272. The ED 50 for this effect is 2.5-15 ng/mL.
Scientific Data Examples for Recombinant Human BMP-4 GMP Protein, CF
Recombinant Human BMP-4 GMP Protein SDS-PAGE
1 μg/lane of GMP-grade Recombinant Human BMP-4 (Catalog # 314-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing bands at 19 - 24 kDa, and 35 - 41 kDa, respectively.
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.
314-GMP
Formulation
Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA.
Reconstitution
Reconstitute at 50-200 μg/mL in 4 mM HCl.
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 6 months when stored at ≤ -20 degreesC as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 3 months, -70 degreesC under sterile conditions after reconstitution.
Reconstitution Calculator
Background: BMP-4
BMP-4 is a TGF-beta superfamily ligand that is widely expressed from early embryogenesis through adulthood. It plays an important role in mesenchyme formation, epidermal determination, suppression of neural induction, the development of multiple organs, and tissue repair (1-5). The human BMP-4 precursor contains a 273 amino acid (aa) propeptide and a 116 aa mature protein (6). Processing of the propeptide by furin or proprotein convertase 6 enables the formation of the mature disulfide-linked homodimeric BMP-4 and facilitates its secretion. Similar intracellular processes may lead to the formation and recreation of BMP4/BMP7 disulfide-linked heterodimer (7-9). Mature human and mouse BMP-4 share 98% aa sequence identity. Human BMP-4 shares 85% aa sequence identity with human BMP-2 and less than 50% with other human BMPs. Compared to BMP-4 homodimers, BMP-4/BMP-7 heterodimers exhibit a greater potency in inducing osteogenic differentiation (9). In Xenopus, the heterodimers can also induce the formation of mesoderm, whereas BMP-4 homodimers only provide ventralizing signals for existing mesoderm (10). BMP-4 signals through tetrameric complexes composed of type I (primarily Activin RIA or BMPR-IA) and type II (primarily Activin RIIA or BMPR-II) receptors (11, 12). The bioavailability of BMP-4 is regulated by its interaction with multiple proteins and glycosaminoglycans (13-15).
Item
Value
References
Zhang, P. et al. (2008) Blood 111:1933.
Gambaro, K. et al. (2006) Cell Death Differ. 13:1075.
Simic, P. and S. Vukicevic (2005) Cytokine Growth Factor Rev. 16:299.
Sadlon, T.J. et al. (2004) Stem Cells 22:457.
Frank, D.B. et al. (2005) Circ. Res. 97:496.
Wozney, J. et al. (1988) Science 242:1528.
Cui, Y. et al. (1998) EMBO J. 17:4735.
Cui, Y. et al. (2001) Genes Dev. 15:2797.
Aono, A. et al. (1995) Biochem. Biophys. Res. Commun. 210:670.
Nishimatsu, S. and G.H. Thomsen (1998) Mech. Dev. 74:75.
Chen, D. et al. (2004) Growth Factors 22:233.
Lavery, K. et al. (2008) J. Biol. Chem. April 24 epub.
Rosen, V. (2006) Ann. N.Y. Acad. Sci. 1068:19.
Jones, C.M. and J.C. Smith (1998) Dev. Biol. 194:12.
Takada, T. et al. (2003) J. Biol. Chem. 278:43229.
BMP-2B, BMP-4, BMP2B, BMP2B1, BMP2BMCOPS6, BMP4, Bone morphogenetic protein 2B, DVR4, MCOPS6, OFC11, ZYME, bone morphogenetic protein 4
Manufacturing Specifications
GMP Proteins R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: WHO TRS, No. 822, 1992 Annex 1, Good Manufacturing Practices for Biological Products; USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and USP Chapter 92, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.
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
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 research. The Certificate of Analysis provided contains the following lot specific information:
N-terminal amino acid analysis, SDS-PAGE analysis, 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
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, 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.
Product Specific Notices for Recombinant Human BMP-4 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.
We suggest you print and retain a copy of these End User Terms of Use of Product for your records.
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.
R&D Systems, Inc. has the right, at its sole discretion, to modify, add or remove any terms or conditions of these End User Terms of Use without notice or liability to you. Any changes to these End User Terms of Use are effective immediately following the printing of such changes on this product insert. The most recent version of these End User Terms of Use of Product may be found at: RnDSystems.com/Legal .
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
The following limitation applies to R&D Systems' warranty and liability for damages: All products are warranted to meet R&D Systems' published specifications when used under normal laboratory conditions.
R&D SYSTEMS DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO ITS PRODUCTS. IN PARTICULAR, R&D SYSTEMS DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NONINFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE.
NOTWITHSTANDING ANY OTHER PROVISIONS OF THESE TERMS AND/OR ANY OTHER AGREEMENT BETWEEN R&D SYSTEMS AND PURCHASER FOR THE PURCHASE OF THE PRODUCTS, R&D SYSTEMS' TOTAL LIABILITY TO PURCHASER ARISING FROM OR IN RELATION TO THESE TERMS, AN AGREEMENT BETWEEN THE PARTIES OR THE PRODUCTS, WHETHER ARISING IN CONTRACT, TORT OR OTHERWISE SHALL BE LIMITED TO THE TOTAL AMOUNT PAID BY PURCHASER TO R&D SYSTEMS FOR THE APPLICABLE PRODUCTS. IN NO EVENT WILL R&D SYSTEMS BE LIABLE FOR THE COST OF PROCUREMENT OF SUBSTITUTE GOODS.
Full details of R&D Systems' Terms and Conditions of Sale can be found online at: RnDSystems.com/Legal .
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