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For quantitative determination of human/rat NOS-I in tissue extract samples

Human/Rat NO Synthase-I[NOS-I] EIA

Background

This enzyme immunoassay (EIA) kit is a stable and convenient assay system for NO synthase-I (NOS-I) in human/rat tissue extract. The EIA kit is prepared by using synthetic NOS-I (998-1024) as standard and biotinylated NOS-I (998-1024) as labeled antigen. The kit contains specific polyclonal antibody to recognize human/rat NOS-I.

Feature and Advantages

This EIA kit is used for quantitative determination of human/rat NOS-I in tissue extract samples. The kit is characterized by its sensitive quantification and high specificity. In addition, it has no influences by other components in samples. Human NOS-I (998-1024) standard of this kit is a highly purified synthetic product (purity: higher than 98%). HPLC purified biotinylated glycylglycyl-human NOS-I (998-1024) is used as labeled antigen.

  • The assay kit can measure human/rat NOS-I in the range of 0.412-100 ng/mL
  • The assay completed within 16-20 hr + 1.5 hr.
  • With one assay kit, 41 samples can be measured in duplicate
  • Test sample: human and rat tissue extract Sample volume: 50 μL
  • The 96-well plate in kit was consisted by 8-wells strips. The kit can be used separately.
  • Precision and reproducibility
    Intra-assay CV(%) : 4.0-5.3
    Inter-assay CV(%) : 4.7-8.0
  • Stability and Storage  Store all of the components at 2-8°C.
    The kit is stable under the condition for 6 months from the date of manufacturing. The expiry date is indicated on the label of the kit.

<Specificity>
The EIA kit shows cross reactivity of 100% to human NOS-I and rat NOS-I

<Test Principle>
This EIA kit for determination of human/rat NOS-I in tissue extract samples is based on a competitive enzyme immunoassay using combination of highly specific antibody to human/rat NOS-I and biotin-avidin affinity system. The 96-wells plate is coated with goat anti rabbit IgG, NOS-I standard or samples, labeled antigen and rabbit anti NOS-I antibody are added to the wells for competitive immunoreaction. After incubation and plate washing, HRP labeled streptoavidin (SA-HRP) is added to form HRP labeled streptoavidin-biotinylated NOS-I-antibody complex on the surface of the wells. Finally, HRP enzyme activity is determined by color reaction of o-phenylenediamine dihydrochloride (OPD) and the concentration of human/rat NOS-I is calculated.

Composition

Component Form Quantity Main Ingredient
1. Antibody coated plate MTP*1 1 plate (96 wells) Goat anti rabbit IgG
2. NOS-I Standard lyophilized 1 vial (100ng) Synthetic human NOS-I (998-1024)
3. Labeled antigen lyophilized 1 vial Biotinylated human NOS-I (998-1024)
4. Specific antibody Liquid 1 bottle (6 mL) Rabbit anti human NOS-I (998-1024) IgG
5. SA-HRP solution liquid 1 bottle (22mL) HRP labeled streptoavidin
6. OPD tablet tablet 2 tablets o-Phenylenediamine dihydrochloride
7. Substrate buffer Liquid 1 bottle (25mL) Citrate buffer containing  0.015% Hydrogen peroxide
8. Stopping solution Liquid 1 bottle (6 mL) 1M H2SO4
9.Washingsolution  (concentrated) Liquid 1 bottle (25 mL) Concentrated saline
10. Buffer solution Liquid 1 bottle (25 mL) Phosphate buffer
11. Adhesive foil   3 pieces  

MTP*1 ….. Microtitration plat

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