Human IGFBP4 ELISA Kit

Catalog No.: BEK1262

Size: 96T

Range: 500 pg/ml - 32000 pg/ml

Sensitivity < 20 pg/ml

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

Application: For quantitative detection of IGFBP4 in human serum, plasma, body fluids, tissue homogenate or cell culture supernatants.

Introduction

Insulin-like growth factor-binding protein 4 is a protein that in humans is encoded by the IGFBP4 gene. IGFBP4 maternal blood concentrations were lower during pregnancy than during non-pregnant cycles and synchronized control cycles. It can be speculated that the lower IGFBP4 in maternal blood may result in an increase of free IGF1 for local action. IGFBP4 downregulation in GCTs compared to MSCs. Overexpression of UCHL1 and IGFBP4 by stable transfection of GCTSC did not influence cell viability, proliferation, migration and chemosensitivity compared to parental cells. However, colony-formation was significantly decreased suggesting that rescue of UCHL1 and IFGBP4 suppresses clonogenicity of GCT stromal cells. Epigenetic silencing of UCHL1 and IGFBP4 in GCTs might thus be a crucial event during the malignant transformation of MSCs in the context of GCT development and represent promising targets for the development of new diagnostic and therapeutic strategies.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- IGFBP4 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- IGFBP4 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 IGFBP4 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of IGFBP4 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-Human IGFBP4 antibody
2. Lyophilized Human IGFBP4 standards: 2 tubes (32 ng / tube)                     
3. Sample / Standard diluent buffer: 30 ml
4. Biotin conjugated anti-Human IGFBP4 antibody (Concentrated): 130 μl. Dilution: 1:100
5. Antibody diluent buffer: 12 ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. 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