Human ETS varient transcription factor 4, ETV4 ELISA Kit

Catalog No.: BZEK2235-48

Size: 48T

Range: 10 pg/ml - 160 pg/ml

Sensitivity: 1 pg/ml

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

Application: For quantitative detection of ETV4 in human serum, plasma, tissue homogenate or cell culture supernatant.

Introduction

ETS translocation variant 4 (ETV4), also known as polyoma enhancer activator 3 (PEA3), is a member of the PEA3 subfamily of Ets transcription factors. Two variants of a disease associated with ETV4 is Ewing Sarcoma and Extraosseous Ewing's Sarcoma. While both are cancerous tumors, the former grows in the bones most commonly affecting the arms, legs, hips, and spine, while the later affects the soft tissue in the chest, foot, pelvis and spine.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- ETV4 antibody was pre-coated onto 48-well plates. And the HRP conjugated anti- ETV4 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 ETV4 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of ETV4 can be calculated.

Kit components

1. One 48-well plate pre-coated with anti- human ETV4 antibody

2. Standard: 0.5ml (320 pg/ml)

3. Standard diluent buffer: 1.5 ml

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

5. Sample diluent buffer: 3 ml

6. HRP conjugated anti- human ETV4 antibody (RTU): 3ml

7. Stop solution: 3 ml

8. TMB substrate A:  3ml

9. TMB substrate B:  3ml

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