Recombinant Human BMP-4 GMP Protein, CF

Catalog #: 314E-GMP Datasheet / COA / SDS

Animal Free

Catalog #	Availability	Size / Price	Qty
314E-GMP-050

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

Recombinant Human BMP-4 GMP Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

Activity

Measured by its ability to induce BMP responsive SEAP reporter activity in HEK293 human embryonic kidney cells. The ED₅₀ for this effect is 3-21 ng/mL.

Source

E. coli-derived human BMP-4 protein
Lys303-Arg408
Produced in an animal-free laboratory. Manufactured and tested under cGMP guidelines

Accession #

P12644.1

N-terminal Sequence
Analysis

Identity was confirmed by mass spectrometry

Structure / Form

Disulfide-linked homodimer

Predicted Molecular Mass

12 kDa

SDS-PAGE

8-9 kDa, reducing conditions

Product Datasheets

314E-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.

314E-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA.
Reconstitution	Reconstitute at 50-200 μg/mL in sterile 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 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, ≤ -65 °C under sterile conditions after reconstitution.

Scientific Data

Bioactivity

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GMP-grade Recombinant Human BMP-4 (Catalog # 314E-GMP) induces BMP responsive SEAP reporter activity in HEK293 human embryonic kidney cells. The ED₅₀ for this effect is 3-21 ng/mL.

Bioactivity

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Equivalent bioactivity of GMP (Catalog # 314E-GMP) and Animal-Free (AFL314E) grades of Recombinant Human BMP-4 as measured by its ability to induce BMP responsive SEAP reporter activity in HEK293 human embryonic kidney cells (orange and green respectively).

SDS-PAGE

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2 μg/lane of GMP-grade Recombinant Human BMP-4 Protein (314E-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 8-9 kDa and 20-22 kDa, respectively.

Reconstitution Calculator

Background: BMP-4

Bone morphogenetic protein 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). It is an integral part of many stem cell differentiation pathways, including lung tissue, (6), adipogenesis (7) and osteogenesis (8, 9). The human BMP-4 precursor contains a 273 amino acid (aa) propeptide and a 116 amino acid (aa) mature protein (10). 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 (11-13). 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. In Xenopus, BMP-4 dimers provide ventralizing signals for existing mesoderm (14). 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 (15, 16). The bioavailability of BMP-4 is regulated by its interaction with multiple proteins and glycosaminoglycans (17-19).

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.
Lee, J-H. et al. (2014) Cell. 156:440.
Tang, QQ. et Lane MD. (2012) Annu Rev. Biochem. 81:715.
Urist, MR. (1965). Science. 150:893.
Bandyopadhyay, A. et al. (2006) Plos Genetics. 2:e216.
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.

Long Name

Bone Morphogenetic Protein 4

Entrez Gene IDs

652 (Human); 12159 (Mouse); 25296 (Rat); 30612 (Zebrafish)

Alternate Names

BMP2B; BMP-2B; BMP2B1; BMP2BMCOPS6; BMP4; BMP-4; Bone morphogenetic protein 2B; bone morphogenetic protein 4; DVR4; MCOPS6; OFC11; ZYME

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:

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 <71>
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 cell therapy applications. 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 in a dedicated cold storage room.

Quality Assurance

Low Endotoxin Level.
No impairment of biological activity.
High quality product obtained under stringent conditions.

Please read our complete Animal-Free Statement.

Product Specific Notices

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.

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Full details of R&D Systems' Terms and Conditions of Sale can be found online at: RnDSystems.com/Legal.