BMP4 ELISA Kit

Catalog No. ABIN1112566

Human BMP4 ELISA Kit Colorimetric assay for quantification of Human BMP4.

Quick Overview for BMP4 ELISA Kit (ABIN1112566)

Target: BMP4 (Bone Morphogenetic Protein 4 (BMP4))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5-4000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma, Serum, Tissue Lysate

Product Details BMP4 ELISA Kit

Minimum Detection Limit: 62.5 pg/mL

Purpose: For quantitative detection of BMP-4 in human serum, plasma, body fluids, tissue lysates or cell culture supernates.

Analytical Method: Quantitative

Sensitivity: < 4 pg/mL

Components: 1. One 96-well plate pre-coated with anti-human BMP-4 antibody 2. Lyophilized BMP-4 standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Human BMP-4 antibody (Concentrated): 130 µl.

Material not included: 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Application Details

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- BMP-4 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- BMP-4 polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the BMP-4 amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of BMP-4 can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Tissue lysate or body fluids, cell culture supernate: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 1000 × g for 10 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with EDTA as the anticoagulant. Centrifuge for 10 min at 1000 x g within 30 min of collection. Analyze immediately or aliquot and store frozen at -20 °C. Heparin or citrate can not be used as anticoagulant here. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Target Details for BMP4

Target: BMP4 (Bone Morphogenetic Protein 4 (BMP4))

Alternative Name: BMP-4

Background: Bone morphogenetic protein 4 (BMP-4) is a member of the transforming growth factor-beta superfamily of regulatory molecules. Bone morphogenetic proteins were originally identified by the ability of demineralized bone extract to induce endochondral osteogenesis in vivo in an extraskeletal site, and known for their ability to induce bone and cartilage development. In human embryonic development, BMP4 is a critical signaling molecule required for the early differentiation of the embryo and establishing of a dorsal-ventral axis. BMP4 is secreted from the dorsal portion of the notochord, and it acts in concert with sonic hedgehog (released from the ventral portion of the notochord) to establish a dorsal-ventral axis for the differentiation of later structures. Increase in expression of BMP4 has been associated with a variety of bone diseases, including the heritable disorder Fibrodysplasia Ossificans Progressiva.

Pathways: Steroid Hormone Mediated Signaling Pathway, Regulation of Muscle Cell Differentiation, Tube Formation, Skeletal Muscle Fiber Development