Human RANK ELISA Kit

Catalog No.: BEK1192

Size: 96T

Range: 62.5 pg/ml-4000 pg/ml

Sensitivity < 2 pg/ml

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

Application: For quantitative detection of RANK in tissue lysates or cell culture supernates. Human serum and plasma should be validated by end users.

Introduction

Receptor Activator of Nuclear Factor κ B (RANK), also known as TRANCE Receptor, is a type I membrane protein. It is expressed on the surface of osteoclasts, dendritic cells and facilitates immune signaling. RANK is involved in their activation upon ligand binding. RANKL (Receptor Activator for Nuclear Factor κ B Ligand) is found on the surface of stromal cells, osteoblasts, and T cells. RANKL and RANK have an essential role in the brain. RANK and RANKL appear to be important regulators of interactions between T cells and dendritic cells.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-RANK polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-RANK 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 RANK amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of RANK can be calculated.

Kit components

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