Human IGFBP6 ELISA Kit

Catalog No.: BEK1263

Size: 96T

Range: 62.5 pg/ml - 4000 pg/ml

Sensitivity < 5 pg/ml

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

Application: For quantitative detection of IGFBP6 in human serum, plasma, body fluids, tissue homogenate or cell culture supernatants.

Introduction

Insulin-like growth factor-binding protein 6 (IGFBP-6) is a protein that in humans is encoded by the IGFBP6 gene. IGFBP6 gene plays an important role in the pathogenesis of breast cancer. Protein IGFBP6 plays an important role in the pathogenesis of many malignant tumors, including breast cancer. Gene IGFBP6 knockdown does not trigger the epithelial-mesenchymal transition in MDA-MB-231 cells, but modifies significantly the expression of many genes involved in this process. A decrease of IGFBP6 expression can involve a decrease in the expression of N-cadherin and transcription factor Slug.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- IGFBP6 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- IGFBP6 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 IGFBP6 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of IGFBP6 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human IGFBP6 antibody
2. Lyophilized Human IGFBP6 standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30 ml
4. Biotin conjugated anti-Human IGFBP6 antibody (Concentrated): 130 μl. Dilution: 1:100
5. Antibody diluent buffer: 12 ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130 μl. Dilution: 1:100
7. ABC diluent buffer: 12 ml
8. TMB substrate: 10 ml
9. Stop solution: 10 ml
10. Wash buffer(25X): 30 ml

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