Fish Luteinizing Hormone, LH ELISA Kit

Catalog No.: BYEK2663

Size: 96T

Range: 0.4 ng/ml - 4.8 ng/ml

Sensitivity: 0.1 ng/ml

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

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

Introduction

Luteinizing hormone (LH, also known as lutropin and sometimes lutrophin is a hormone produced by gonadotropic cells in the anterior pituitary gland. The production of LH is regulated by gonadotropin-releasing hormone (GnRH) from the hypothalamus. In females, an acute rise of LH ("LH surge") triggers ovulation[3] and development of the corpus luteum. In males, where LH had also been called interstitial cell–stimulating hormone (ICSH),[4] it stimulates Leydig cell production of testosterone. It acts synergistically with follicle-stimulating hormone (FSH). LH is a heterodimeric glycoprotein. Each monomeric unit is a glycoprotein molecule; one alpha and one beta subunit make the full, functional protein.

Principle of the Assay  

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

Kit components

1. One 96-well plate pre-coated with anti- fish LH antibody

2. Standard: 0.5ml (7.2 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- fish LH 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