Mouse BMP-4 ELISA Kit

Catalog No.: BEK1280

Size: 96T

Range: 31.2 pg/ml - 2,000 pg/ml

Sensitivity < 4 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months.

Application: For quantitative detection of BMP-4 in mouse serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Bone morphogenetic protein 4 is a protein that in humans is encoded by BMP4 gene. BMP4 is found on chromosome 14q22-q23. BMP4 is a member of the bone morphogenetic protein family which is part of the transforming growth factor-beta superfamily. The superfamily includes large families of growth and differentiation factors. BMP4 is highly conserved evolutionarily. BMP4 is found in early embryonic development in the ventral marginal zone and in the eye, heart blood and otic vesicle.

Principle of the Assay

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 to 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 to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the BMP-4 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration  ofBMP-4 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-mouse BMP-4 antibody
2. Lyophilized mouse BMP-4 standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-mouse BMP-4 antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ 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