Anti-Human BMP-2

Product Datasheet

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Anti-Human Bone Morphogenetic Protein 2 (BMP-2)

Purified No Carrier Protein

(BMP-2A, OP-1)

Monoclonal Antibody

Prod. No.:	B576
Clone:	65529
Isotype:	Mouse IgG_2b
Pkg. Size:	1 mg, 500 µg
Storage:	-20°C to -70°C Detailed storage instructions below.

Description

Specificity:

Mouse Anti-Human Bone Morphogenetic Protein 2 (BMP-2) (Clone 65529) recognizes an epitope on Human BMP-2. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.

Background:

BMPs (Bone Morphogenetic Proteins) belong to the TGF-beta superfamily of structurally related signaling proteins. BMP-2 is a potent osteoinductive cytokine, capable of inducing bone and cartilage formation in association with osteoconductive carriers such as collagen and synthetic hydroxyapatite. In addition to its osteogenic activity, BMP-2 plays an important role in cardiac morphogenesis. The functional form of BMP-2 is a 26 kDa protein composed of two identical 114 amino acid polypeptide chains linked by a single disulfide bond. Each BMP-2 monomer is expressed as the C-terminal part of a precursor polypeptide, which also contains a 23 amino acid signal sequence for secretion, and a 259 amino acid propeptide. After dimerization of this precursor, the covalent bonds between the propeptide (which is also a disulfide-linked homodimer) and the mature BMP-2 ligand are cleaved by a furin-type protease.

Human and mouse BMP-2 and BMP-7 are 100% and 98% identical, respectively, at the amino acid level. Human BMP-2 shares 85% aa sequence identity with human BMP-4 and less than 51% aa sequence identity with other BMPs. Human BMP-7 shares approximately 60% - 70% aa sequence identity with BMP-5, -6, and -8, and less than 50% aa sequence identity with other BMPs. BMP-2 and BMP-7 are co-expressed in some embryonic tissues 6,7 and associate into a functional 38 kDa osteogenic dimer.8 In in vitro osteoblast differentiation assays and in vivo bone formation models, a BMP-2/BMP-7 heterodimer is significantly more potent than either homodimer.

Bone morphogenetic protein 2 is shown to stimulate the production of bone and recombinant human protein (rhBMP-2) and is currently available for orthopaedic usage in the United States. Implantation of BMP-2 in a collagen sponge induces new bone formation and can be used for the treatment of bony defects, delayed union, and non-union. Bone morphogenetic protein 2 has also found its way into the field of Dentistry. Oral Surgery and Implant Dentistry in particular have benefited dramatically from commercially available BMP-2.

Known Species Cross Reactivity: Hu+

Host Species

Mouse

Immunogen

Purified Recombinant Human BMP-2 (>98%)

Formulation

This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.

Reconstitution

This lyophilized antibody may be aseptically reconstituted with sterile PBS to a final volume of 1.0 ml. This will result in a final antibody concentration of 500 µg/ml.

Storage and Stability

The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.

Reported Applications

Indirect ELISA, WB

Indirect ELISA: This antibody can be used in an indirect detection ELISA at 0.5 - 1.0 µg/ml with a suitable second step reagent such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114). The detection sensitivity of this indirect ELISA for Human BMP-2 is approximately 12 ng/well.

Western Blotting: To detect Human BMP-2 this monoclonal antibody can be used at a concentration of 1-2 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The detection limit for Human BMP-2 is 25 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. A suitable Western blotting control is Recombinant Human BMP-2 (Leinco Prod. No.: B388).

Each investigator should determine their own optimal working dilution for specific applications.

Country of Origin

USA