Human Vimentin ELISA Kit

Catalog No.: BEK1237

Size: 96T

Range: 20 ng/ml-800 ng/ml

Storage and Expiration: Store at 2-8℃ for 6 months. 

Application: For quantitative detection of Vimentin in Human serum, plasma, urine, cell culture supernatant or tissue samples.

Introduction

Vimentin is a type III intermediate filament (IF) protein that is expressed in mesenchymal cells. IF proteins are found in all metazoan cells as well as bacteria. All IF proteins are expressed in a highly developmentally-regulated fashion; vimentin is the major cytoskeletal component of mesenchymal cells. Because of this, vimentin is often used as a marker of mesenchymally-derived cells or cells undergoing an epithelial-to-mesenchymal transition (EMT) during both normal development and metastatic progression. It plays a significant role in supporting and anchoring the position of the organelles in the cytosol, and it has been used as a sarcoma tumor marker to identify mesenchyme.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti-Vimentin antibody was pre-coated onto 96-well plates. And the HRP conjugated anti-Vimentin antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) 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 Vimentin amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of Vimentin can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human Vimentin antibody

2. Standard:  0.5ml (900 ng /mL)

3. Standard diluent buffer:  1.5 ml

4. Wash buffer (30×):  20 ml. Dilution: 1:30

5. Sample diluent buffer: 6 ml

6. HRP conjugated anti-human Vimentin antibody (RTU): 6 ml

7. Stop solution: 6 ml

8. TMB substrate A:  6 ml

9. TMB substrate B:  6 ml

10. Plate sealer:   2

11. Hermetic bag:  1

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. Absorbent filter papers
8. Plastic or glass container with volume of above 1L