Human Desmin ELISA Kit

Catalog No.: BEK1235

Size: 96T

Range: 10 ng/ml-450 ng/ml

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

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

Introduction

Desmin is a type III intermediate filament found near the Z line in sarcomeres. It was first described in 1976, first purified in 1977, the gene was cloned in 1989, and the first knock-out mouse was created in 1996. It is one of the earliest myogenic markers, both in heart and somites, and is expressed in satellite stem cells and replicating myoblasts. The desmin provides attachments between the terminal Z disc and membrane-associated proteins to form a force-transmitting system that parallels the thin filaments at myotendinous junctions. Desmin is also important in muscle cell architecture and structure since it connects many components of the cytoplasm. Finally, desmin may be important in mitochondria function. When desmin is not functioning properly there is improper mitochondrial distribution, number, morphology and function.

Principle of the Assay  

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

Kit components

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

2. Standard: 0.5ml (540 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 Desmin antibody (RTU): 6ml

7. Stop solution: 6 ml

8. TMB substrate A:  6ml

9. TMB substrate B:  6ml

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