Mouse NGF / NGFβ ELISA Kit

Catalog No.: BEK1170

Size: 96T

Range: 31.2 pg/ml-2000 pg/ml

Sensitivity < 1 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of NGF in mouse serum, body fluids, tissue lysates or cell culture supernatants.

Introduction

Nerve growth factor (NGF) is a polypeptide involved in the regulation of growth and differentiation of sympathetic and certain sensory neurons. NGF consists of 3 types of subunits, alpha, beta and gamma, which specifically interact to form a 7S, approximately 130,000-molecular weight complex. It is mainly expressed in Sertoli and interstitial cells. The proforms of NGF are secreted and cleaved extracellularly by the serine protease plasmin and by selective matrix metalloproteinases. NGF is important for the growth, maintenance, and survival of certain target neurons (nerve cells). It also functions as a signaling molecule.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-NGF polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-NGF polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates 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 NGF amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of NGF can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-mouse NGF antibody
2. Lyophilized Mouse NGF standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Mouse NGF antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

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. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L