Rabbit Nitric Oxide Synthase (NOS) Quantitative Detection Kit (ELISA)

user's Guide

[Rabbit Nitric Oxide Synthase (NOS) Elisa Kit Kit Name]

Rabbit Nitric Oxide Synthase (NOS) Quantitative Detection Kit (ELISA)

[Rabbit Nitric Oxide Synthase (NOS) Elisa Kit Kit Use]

Quantitative detection of nitric oxide synthase (NOS) in rabbit serum, plasma and related liquid samples.

[Rabbit Nitric Oxide Synthase (NOS) Elisa kit detection principle]

This kit uses double-antibody two-step sandwich enzyme-linked immunosorbent assay (ELISA). Add the standard and the test sample to the pre-coated rabbit nitric oxide synthase (NOS) monoclonal antibody transparent enzyme label coated plate. After incubation for a sufficient time, wash to remove unbound components, and then add the enzyme label to work After incubation for a sufficient period of time, wash to remove unbound components. Substrates A and B are added in sequence, and the substrate (TMB) is converted into a blue product catalyzed by horseradish peroxidase (HRP) and turns yellow under the action of an acid. Synthetase (NOS) concentration was positively correlated. The OD value was measured at 450 nm wavelength. Based on the OD value of the standard and the sample, the rabbit nitric oxide synthase (NOS) content was calculated.

[Composition of Rabbit Nitric Oxide Synthase (NOS) Elisa Kit Kit]

1

Enzyme coated plate

12 holes × 8

7

Developer A liquid

6mL

2

Standard

0.3mL × 6 tubes

8

Developer B liquid

6mL

3

20 times concentrated washing liquid

25mL

9

Stop solution

6mL

4

Sample diluent

6mL

10

Instructions

1 serving

5

Special diluent

6mL

11

Sealing film

2 sheets

6

Enzyme reagent

6mL

12

sealed bag

1

Remarks: The concentration of standard products (No. 1 → No. 6) is: 48, 24, 12, 6, 3, 1.5 μmol / L.

[Reagents and equipment not required by rabbit nitric oxide synthase (NOS) Elisa kit]

1. 37 ℃ thermostat

2. Standard specification microplate reader

3. Precision pipettes and disposable tips

4. Distilled water

5. Disposable test tubes

6. Absorbent paper

[Operation steps of rabbit nitric oxide synthase (NOS) Elisa kit]

1. Preparation: Remove the reagent kit from the refrigerator and re-equilibrate at room temperature for 30 minutes.

2. Mixing solution: dilute the 20-fold concentrated washing solution with distilled water to the original one.

3. Add standard products and samples to be tested: take a sufficient number of enzyme-coated plates and fix them to the frame. Set up standard wells, sample wells to be tested and blank control wells, record the positions of each well in the standard wells. Add 50μL of standard product; first add 10μL of sample to be tested, and then add 40μL of sample diluent (that is, the sample is diluted 5 times); blank control well is not added.

4. Incubation: Incubate in a 37 ° C water bath or thermostat for 30 minutes.

5. Wash the plate: discard the liquid, pat dry on the absorbent paper, fill each hole with the washing solution, let stand for 1min, shake off the washing solution, pat dry on the absorbent paper, repeat the washing 4 times (you can also use the washing machine to press Instructions for washing the board).

6. Add enzyme-labeled working solution: add 50μL of enzyme-labeled working solution to each well, without adding blank control wells.

7. Incubation: Incubate in a 37 ° C water bath or thermostat for 30 minutes.

8. Wash the plate: discard the liquid, pat dry on the absorbent paper, fill each hole with washing liquid, let stand for 1min, shake off the washing liquid, pat dry on the absorbent paper, repeat the plate washing 4 times (you can also use the washing machine to press Instructions for washing the board).

9. Color development: add 50μL of developer A solution to each well, and then add 50μL of developer B solution, mix for 30s with a plate mixer (or gently shake for 30s by hand), and avoid color development at 37 ℃ for 15min .

10. Termination: Remove the enzyme labeling plate and add 50μL of stop solution to each well to stop the reaction (the color changes from blue to yellow).

11. Determination: Zero the blank holes, and within 15 minutes after termination, measure the absorbance (OD value) of each well with a wavelength of 450 nm.

12. Calculation: According to the concentration of the standard product and the corresponding OD value, calculate the linear regression equation of the standard curve, and then calculate the corresponding sample concentration on the regression equation according to the OD value of the sample. You can also use various application software to Calculation. The final concentration is the actual measured concentration times the dilution factor.

[Rabbit nitric oxide synthase (NOS) Elisa kit sample requirements]

1. The sample cannot contain sodium azide (NaN3) because sodium azide (NaN3) is an inhibitor of horseradish peroxidase (HRP).

2. As soon as possible after specimen collection

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