BMP5 antibody (ab10858)

Product Name 

BMP5 antibody

Product type 

Primary antibodies

Description 

Goat polyclonal to BMP5

Immunogen 

Recombinant full length protein (Human) expressed in NSO cells.

Reacts with 
(species key)

Hu

Specificity 

Based on ELISA, this antibody shows less than 5 % cross-reactivity with recombinant human BMP2 and recombinant human BMP4.

Tested applications 
(see key)

ELISA, IHC-Fr, IHC-P, Inhib, WB

Abreviews 

Customer reviews (see key)

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Application notes 
(see key)

Recommended dilutions
ELISA: Use at an assay dependent dilution. The detection limit for recombinant human BMP5 is approximately 0.16 ng/well.
IHC-Fr: Use at a concentration of 1 - 3 µg/ml.
IHC-P: Use at a concentration of 5 - 10 µg/ml.
Inhib: Use at an assay dependent dilution.
WB: Use at an assay dependent dilution. Predicted molecular weight: 20 kDa.

Cells may be fixed for 20 minutes at room temperature with freshlyprepared 1 to 2 % paraformaldehyde/PBS (pH 7.4). Three to five washes of the cells in PBS (15 minutes each) is usually required after fixation and before the addition of the primary antibody (anti-BMP5). Labeling may be obtained by incubating the cells overnight at 4 °C with at least 1 to 3 µg/ml anti-BMP5 antibody followed by the appropriate secondary reagents.

For tissue immunohistochemistry, a working concentration of at least 5 to 10 µg/ml antibody is recommended to detect human BMP5 on cryostat sections 5 to 15 µm thick. Dissected tissues should be fixed by vascular perfusion with 4 % paraformaldehyde/ PBS (pH 7.4) and followed by perfusion with a 10 % sucrose solution in 0.1 M phosphate buffer (pH 7.2). Note: prolonged fixation in 4 % paraformaldehyde can diminish the labeling and staining intensity of BMP5. Cryostat sections from 5 to 15 µm are incubated with the primary antibody (anti-BMP5) at concentrations of at least 5 to 10 µg/ml. Note: on freefloating sections, anti-BMP5 should be diluted to at least 0.3 to 1 µg/ml.

The labeling detection system on cells and tissues can be done by either fluorescent or non-fluorescent enzymatic assays. Due to autofluorescence in neuronal tissues, the use of fluorescence detectors such as FITC or Cy3 is not recommended unless the autofluorescence is quenched (example: treating tissues after IHC staining with 1% Sudan Black in 70 % alcohol). Non-fluorescent enzymatic staining using DAB, AEC, or immunogold-silver can be used.

Anti-human BMP5 has the ability to neutralize the biological activity of recombinant human BMP5 on MC3T3-E1 cells. Recombinant human BMP5 is added to various concentrations of the antibody for 1 hour at 37 °C in a 96 well microplate. Following this pre-incubation, MC3T3-E1 cells are added to the mixture. The assay mixture in a total volume of 100 µl, containing antibody at concentrations of 0.01 µg/ml to 100 µg/ml, recombinant human BMP5 at 1.5 µg/ml, L-ascorbic acid at 50 µg/ml, and cells at 5 x 10⁴ cells/ml, is incubated at 37 °C for 4 days in a humidified CO2 incubator. At the end of the incubation, alkaline phosphatase activity in cell lysate is measured.

The Neutralization Dose50 (ND50) for anti-human BMP5 is approximately 3 to 10 µg/ml in the presence of 1.5 µg/ml of recombinant human BMP5, using the MC3T3-E1 cell line. The ND50 is the concentration of antibody required to yield one-half maximal inhibition of the cytokine activity on a responsive cell line, when the cytokine is present at a concentration just high enough to elicit a maximum response. The exact concentration of antibody required to neutralize human BMP5 activity is dependent on the cytokine concentration, cell type, growth conditions, and the type of activity studied.

The detection limit for recombinant human BMP5 is approximately 5 ng/lane under non-reducing and reducing conditions. Recombinant BMP5, a disulfide-linked homodimeric protein, consists of two 167 amino acid residue subunits with a calculated molecular mass of approximately 18 kDa. Due to glycosylation, the protein migrates as a doublet of 20 kDa and 25 kDa under reducing conditions in SDS-PAGE. BMP5 is synthesized as a large precursor protein that is cleaved at the dibasic cleavage site (RXXR) to release the carboxy-terminal domain.

Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.

Cellular localization 

Secreted

Research areas 

Stem Cells >> Signaling Pathways >> TGF beta >> Secreted
Signal Transduction >> Cytoskeleton / ECM >> Extracellular Matrix >> Structures >> Bone
Signal Transduction >> Growth Factors/Hormones >> TGF
Signal Transduction >> Cytoskeleton / ECM >> Extracellular Matrix >> ECM Proteins >> Collagen
Cardiovascular >> Angiogenesis >> Growth Factors >> TGF

Relevance 

Bone Morphogenetic Proteins (BMPs) are members of the TGF beta superfamily of cytokines that affect bone and cartilage formation. Similar to other TGF beta family proteins, BMPs are highly conserved across animal species. Mature BMPs are 30-38 kDa proteins that assume a TGF beta like cysteine knot configuration. Unlike TGF beta, BMPs do not form latent complexes with their propeptide counterparts. Most BMPs are homodimers, but bioactive natural heterodimers have been reported. Recently it was found that lovostatin, widely used for lowering cholesterol, also increases bone formation by turning on a gene (bmp2) that promotes local bone formation. BMPs create an environment conducive for bone marrow development by stimulating the production of specific bone matrix proteins and altering stromal cell and osteoclast proliferation. In addition to stimulating ectopic bone and cartilage development, BMPs may be an important factor in the development of the viscera, with roles in cell proliferation, apoptosis, differentiation, and morphogenesis. BMPs also appear to be responsible for normal dorsal/ventral patterning. BMPs are found in tissues that induce bone or cartilage growth, such as demineralized bone and urinary epithelium. Cellular responses to BMP5 are mediated by the formation of hetero-oligomeric complexes of type I and type II serine/threonine kinase receptors which play significant roles in BMP binding and signaling. One BMP type II receptor and two BMP type I receptors have been identified.
BMP5 induces cartilage and bone formation and is expressed in lung and liver tissue.

Database links 

Raised in 

Goat

Clonality 

Polyclonal

Isotype 

IgG

Purity 

Immunogen affinity purified

Storage buffer 

Preservative: None
Constituents: PBS

Form 

Liquid

Concentration 

0.100 mg/ml

Storage instructions 

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

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