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

Biotin

(BMP-2A, OP-1)

Polyclonal Antibody

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Prod. No.:	B610
Host:	Goat
Pkg. Size:	50 µg, 1 mg
Storage:	-20°C to -70°C Detailed storage instructions below.

Description

Specificity:

Goat Anti-Human Bone Morphogenetic Protein 2 (BMP-2) recognizes Human BMP-2 and Human BMP-4. This antigen affinity purified polyclonal antibody was purified using a proprietary chromatographic technique that includes covalently immobilizing the antigen proteins or peptides to agarose based beads. This purification method enhances specificity, reduces nonspecific binding of extraneous IgG and provides you with the most reliable reagent available for your early discovery research.

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+

Immunogen

Purified Recombinant Human BMP-2 (>98%)

Formulation

This biotinylated antigen affinity purified polyclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco’s phosphate buffered saline (1X PBS) pH 7.2 – 7.3 containing 50 µg of bovine serum albumin per µg of antibody with no calcium, magnesium, or preservatives present.

Reconstitution

We suggest aseptically reconstituting this biotinylated, lyophilized, antigen affinity purified polyclonal antibody with 1 ml of sterile 20 mM Trizma base, 0.15 M sodium chloride (1X Tris Buffered Saline) pH 7.2-7.3, containing 0.1% w/v bovine serum albumin (BSA). This will result in a final biotinylated antibody concentration of 50 µg/ml.

Storage and Stability

The lyophilized, biotinylated antigen affinity purified polyclonal antibody can be stored desiccated at -20°C to -70°C for up to twelve months from date of receipt. The reconstituted bioin conjugate can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted conjugate, 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

IHC (NBF/Par.), WB

IHC (NBF/Par.): This antibody should give satisfactory staining results when used at a concentration of 10 µg/ml. The recommended secondary antibody for IHC is UltraAvidin™- Horseradish Peroxidase (HRPO) (Leinco Prod. No.: A106). For chromogenic detection with high signal and low background use 3, 3'-Diaminobenzidine Tetrahydrochloride (DAB) Substrate Kit (Leinco Prod. No.: D100) or Optional Metal Enhanced DAB Substrate Kit (Leinco Prod. No.: K107).

Western Blotting: To detect Human BMP-2 this biotin conjugate can be used at a concentration of 0.1 - 0.2 µg/ml. This biotin conjugate should be used in conjunction with compatible second-step reagents such as UltraAvidin™- Horseradish Peroxidase (HRPO) (Leinco Prod. No.: A106) 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.: B625).

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

Country of Origin

USA