Rat NT-4 ELISA Kit

Catalog No.: BEK1176

Size: 96T

Range: 600 pg/ml-20,000 pg/ml

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

Application: For quantitative detection of NT-4 in rat serum, plasma, urine, cell culture supernatant or tissue samples.

Introduction

Neurotrophins act as survival and differentiation factors in the nervous system and have been detected in the developing rodent testis. Three additional structurally homologous neurotrophic factors have been identified. These include brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4 (NT-4) (also designated NT-5). Neurotrophin-4 (NT-4), is a neurotrophic factor that signals predominantly through the TrkB receptor tyrosine kinase. Mammalian NT4 had many unusual features compared to the previously identified neurotrophins and was less conserved evolutionarily than the others; however, mammalian NT4 displayed bioactivity and TRK receptor specificity similar to that of Xenopus NT4.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti-NT-4 antibody was pre-coated onto 96-well plates. And the HRP conjugated anti-NT-4 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 NT-4 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of NT-4 can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-rat NT-4 antibody

2. Standard: 0.5ml (22500 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 NT-4 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