Rat NOS1/nNOS (Nitric Oxide Synthase 1, Neuronal) CLIA Kit-Laboratory Biological Research Reagents Manufacturer for Life Science - Elabscience

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Rat NOS1/nNOS (Nitric Oxide Synthase 1, Neuronal) CLIA Kit

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Price:   $682
Cat Number:  E-CL-R0746
Lead Time:   Order now, ship in 3 days

Reactivity:  Rat

Detection Range:  31.25~2000 pg/mL

Sensitivity:  18.75 pg/mL

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Intended use

This CLIA kit applies to the in vitro quantitative determination of Rat NOS1/nNOS concentrations in serum, plasma and other biological fluids.

Test principle

This CLIA kit uses the Sandwich-CLIA principle. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Rat NOS1/nNOS. Standards or samples are added to the micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat NOS1/nNOS and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat NOS1/nNOS, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Rat NOS1/nNOS. You can calculate the concentration of Rat NOS1/nNOS in the samples by comparing the RLU value of the samples to the standard curve.

Assay type Sandwich-CLIA
Format 96T
Assay time 3.5h
Reactivity Rat
Detection method Chemiluminescence
Detection range 31.25-2000 pg/mL
Sensitivity 18.75 pg/mL
Sample volume 100μL
Sample type serum, plasma and other biological fluids
Repeatability CV < 15%

Specificity

This kit recognizes Rat NOS1/nNOS in samples. No significant cross-reactivity or interference between Rat NOS1/nNOS and analogues was observed.

Typical data

As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(pg/mL)
RLU Average Corrected
2000 51196
60246
55721 55693
1000 21612
26336
23974 23946
500 11626
10438
11032 11004
250 5104
5484
5294 5266
125 2648
2568
2608 2580
62.5 1335
1287
1311 1283
31.25 653
695
674 646
0 27
29
28 --
CLIA-Elabscience

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat NOS1/nNOS were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat NOS1/nNOS were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 97.20 307.05 752.28 95.17 326.18 819.78
Standard deviation 12.07 30.09 61.31 9.53 30.01 77.39
C V (%) 12.42 9.80 8.15 10.01 9.20 9.44

Recovery

The recovery of Rat NOS1/nNOS spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5) 87-98 93
EDTA plasma (n=5) 86-98 93
Cell culture media (n=5) 89-105 96

Linearity

Samples were spiked with high concentrations of Rat NOS1/nNOS and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

    Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2 Range (%) 95-110 88-101 95-106
Average (%) 101 93 100
1:4 Range (%) 88-102 86-99 90-104
Average (%) 96 94 97
1:8 Range (%) 90-105 94-107 99-111
Average (%) 97 100 105
1:16 Range (%) 96-107 98-111 94-109
Average (%) 101 104 101

Kit Components& Storage

An unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro CLIA Plate(Dismountable) 8 wells ×12 strips -20℃, 6 months
Reference Standard 2 vials
Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20℃(shading light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 4℃, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent A 1 vial, 5 mL 4℃ (shading light)
Substrate Reagent B 1 vial, 5 mL 4℃ (shading light)
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy

Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

Other supplies required

  • Chemiluminescence immunoassay analyzer
  • High-precision transfer pipette, EP tubes and disposable pipette tips
  • Incubator capable of maintaining 37℃
  • Deionized or distilled water
  • Absorbent paper
  • Loading slot for Wash Buffer

Assay procedure

1.Add 100 μL of standard or sample to each well. Incubate for 90 min at 37℃.

2.Remove the liquid.

3.Add 100 μL Biotinylated Detection Ab. Incubate 1 hour at 37℃. Aspirate and wash 3 times.

4.Add 100 μL HRP Conjugate. Incubate 30 min at 37℃. Aspirate and wash 5 times.

5.Add 100 μL Substrate Mixture Solution. Incubate for 5 min at 37℃.

6.Fluorescence appeared. Measure the RLU value with the Chemiluminescence immunoassay analyzer. Calculation of the results.

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