Human Erythroferrone, ERFE ELISA Kit

Catalog No.: BZEK2268

Size: 96T

Range: 30 pg/ml - 480 pg/ml

Sensitivity: 3 pg/ml

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

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

Introduction

Erythroferrone is a protein hormone encoded in humans by the ERFE gene. Erythroferrone is produced by erythroblasts, inhibits the production of hepcidin in the liver, and so increases the amount of iron available for hemoglobin synthesis. Skeletal muscle secreted ERFE has been shown to maintain systemic metabolic homeostasis. Erythroferrone in humans is transcribed as a precursor of 354 amino acids, with a signal peptide of 28 amino acids. The mouse gene encodes a 340 amino acid protein which is 71% identical. Homology is greater at the C-terminal where there is a TNF-alpha-like domain. As a member of the C1q/TNF-Related Protein (CTRP) family, erythroferrone has a 4-domain structure with a unique N-terminus. The two larger domains are connected by a short, proline-rich, collagenous linker that is thought to promote protein multimerization. Erythroferrone is predicted to contain two PCSK3/furin recognition sites.

Principle of the Assay  

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

Kit components

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

2. Standard: 0.5ml (960 pg/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 ERFE 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