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:

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