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Recombinant Human BMP-4 Animal-Free Protein

R&D Systems, part of Bio-Techne | Catalog # AFL314E

Intended for preclinical researchers who may transition to GMP BMP-4 for their clinical work
R&D Systems, part of Bio-Techne
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AFL314E-010
AFL314E-01M
AFL314E-050
AFL314E-250

Key Product Details

Source

E. coli

Accession #

Structure / Form

Disulfide-linked homodimer

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

E. coli-derived human BMP-4 protein
Lys303-Arg408
Produced using non-animal reagents in an animal-free laboratory.

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.

N-terminal Sequence Analysis

Lys303

Predicted Molecular Mass

12 kDa

SDS-PAGE

8-9 kDa, under reducing conditions

Activity

Measured by its ability to induce BMP responsive SEAP reporter activity in HEK293 human embryonic kidney cells.
The ED50 for this effect is 3.00-21.0 ng/mL.

Scientific Data Images for Recombinant Human BMP-4 Animal-Free Protein

Recombinant Human BMP-4 Animal-Free Protein Bioactivity

Recombinant Human BMP-4 Animal-Free Protein Bioactivity

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.

Equivalent Bioactivity of GMP, Animal-Free, and RUO grades of Recombinant Human BMP-4.

Equivalent bioactivity of GMP (314-GMP), Animal-Free (Catalog # AFL314E) and RUO (314-BPE) 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, green, red, respectively).
Recombinant Human BMP-4 Animal-Free Protein SDS-PAGE

Recombinant Human BMP-4 Animal-Free Protein SDS-PAGE

2 μg/lane of Animal-FreeTM Recombinant Human BMP-4 Protein (AFL314E) 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.

Formulation, Preparation and Storage

AFL314E
Formulation Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA.
Reconstitution Reconstitute at 100-200 μg/mL in 4 mM HCl.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -70 °C under sterile conditions after reconstitution.

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

  1. Zhang, P. et al. (2008) Blood 111:1933.
  2. Gambaro, K. et al. (2006) Cell Death Differ. 13:1075.
  3. Simic, P. and S. Vukicevic (2005) Cytokine Growth Factor Rev. 16:299.
  4. Sadlon, T.J. et al. (2004) Stem Cells 22:457.
  5. Frank, D.B. et al. (2005) Circ. Res. 97:496.
  6. Lee, J-H. et al. (2014) Cell. 156:440.
  7. Tang, QQ. et Lane MD. (2012) Annu Rev. Biochem. 81:715.
  8. Urist, MR. (1965). Science. 150:893.
  9. Bandyopadhyay, A. et al. (2006) Plos Genetics. 2:e216.
  10. Wozney, J. et al. (1988) Science 242:1528.
  11. Cui, Y. et al. (1998) EMBO J. 17:4735.
  12. Cui, Y. et al. (2001) Genes Dev. 15:2797.
  13. Aono, A. et al. (1995) Biochem. Biophys. Res. Commun. 210:670.
  14. Nishimatsu, S. and G.H. Thomsen (1998) Mech. Dev. 74:75.
  15. Chen, D. et al. (2004) Growth Factors 22:233.
  16. Lavery, K. et al. (2008) J. Biol. Chem. April 24 epub.
  17. Rosen, V. (2006) Ann. N.Y. Acad. Sci. 1068:19.
  18. Jones, C.M. and J.C. Smith (1998) Dev. Biol. 194:12.
  19. Takada, T. et al. (2003) J. Biol. Chem. 278:43229.

Long Name

Bone Morphogenetic Protein 4

Alternate Names

BMP2B, BMP2B1, BMP4, MCOPS6, OFC11

Entrez Gene IDs

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

Gene Symbol

BMP4

UniProt

Additional BMP-4 Products

Product Documents for Recombinant Human BMP-4 Animal-Free Protein

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Manufacturing Specifications

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.
    • For ex vivo research or bioproduction, additional documentation can be provided. 

    Please read our complete Animal-Free Statement

    Product Specific Notices for Recombinant Human BMP-4 Animal-Free Protein

    For research use only

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