Recombinant human/mouse/rat/bovine/porcine BMP-2 protein

Summary

• High purity human bone morphogenetic protein 2, BMP-2 (Uniprot: P12643).
• >98%, by SDS-PAGE quantitative densitometry
• 26 kDa (dimer)
• Expressed in E. coli
• Animal origin-free (AOF) and carrier protein-free.
• Manufactured in our Cambridge, UK laboratories
• Lyophilized from acetonitrile, TFA
• Resuspend in 10 mM HCl at >100 µg/ml (provided with protein and free of charge), prepare single use aliquots, add carrier protein if desired and store frozen at -20°C or -80°C

Featured applications

• Differentiation of human pluripotent stem cells towards extra-embryonic endoderm, mesenchymal, neural lineages, and chondrocytes

Purity

BMP-2 migrates as a single band at 26 kDa in non-reducing (NR) conditions and 13 kDa as a single monomeric species upon reduction (R). No contaminating protein bands are visible. Purified recombinant protein (7 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced conditions (NR) and stained with Coomassie Brilliant Blue R250. Data from Qk007 lot #010

R-spondin 1 (Qk006) has be tested at 100 ng/ml – 500 ng/ml, we would recommend using 100 ng/ml – 1 µg/ml for initial testing and optimisation with culture. It is highly recommended to do a dose response to optimize concentration due to varying culture conditions across different labs.

Qkine BMP-2 is more biologically active than a comparable alternative supplier protein

Bioactivity was determined using a BMP-2-responsive firefly luciferase reporter assay in stably transfected HEK293T cells. Cells were treated with a serial dilution of Qkine BMP-2 (Qk007, green) or alternative BMP-2 (supplier B, black). Firefly luciferase activity was measured and normalized to the control Renilla luciferase activity. Data from Qk007 lot #204510.

Protein background

Bone morphogenetic protein 2 (BMP-2) is a member of the BMP subgroup of the transforming growth factor beta (TGF-β) superfamily. It plays numerous roles in the developing embryo, such as embryonic patterning along the dorso-ventral axis, organogenesis, limb bud formation, and bone and cartilage growth. BMP-2 protein is a potent differentiation factor and directs human pluripotent stem cells towards extra-embryonic endoderm, mesenchymal and neural lineages, and chondrocytes [1] .  Recombinant BMP-2 protein induces bone and cartilage formation in vitro and chondrogenesis in human adult mesenchymal stem cells [2].

Human bone morphogenetic protein 2 (BMP-2) is synthesized as a preproprotein consisting of N-terminal signal peptide, 259 amino acid residue pro-domain and 114 residue mature domain. Proteolytic removal of the propeptide enables mature BMP-2 protein to form active disulfide-linked homodimers, and heterodimers with BMP7.

Mature human BMP-2 protein shares 100% amino acid sequence identity with mouse and rat BMP-2. It also shares 85% amino acid sequence identity with the related protein, BMP-4, and less than 51% identity with other BMPs.

Background references