Rat Nuclear Respiratory Factor 1, NRF1 ELISA Kit

Catalog No.: BZEK2382

Size: 96T

Range: 25 pg/ml - 400 pg/ml

Sensitivity: 2 pg/ml

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

Application: For quantitative detection of NRF1 in rat serum, plasma, tissue homogenate or cell culture supernatant.

Introduction

Nuclear respiratory factor 1, also known as Nrf1, Nrf-1, NRF1 and NRF-1, encodes a protein that homodimerizes and functions as a transcription factor which activates the expression of some key metabolic genes regulating cellular growth and nuclear genes required for respiration, heme biosynthesis, and mitochondrial DNA transcription and replication. The Nrf proteins are important for the upregulation of antioxidant and xenobiotic-metabolizing enzymes during oxidative stress. Nrf1 is also involved in heme biosynthesis through regulation of globin gene expression. The protein has also been associated with the regulation of neurite outgrowth.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- NRF1 antibody was pre-coated onto 96-well plates. And the HRP conjugated anti- NRF1 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 NRF1 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of NRF1 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti- rat NRF1 antibody

2. Standard: 0.5ml (800 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- rat NRF1 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