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3-NT(3-Nitrotyrosine) ELISA Kit

  • Cat.No.:E-EL-0040

  • Reactivity: Universal

To Purchase E-EL-0040

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $495
Qty:

Product Details

Properties

Assay type Competitive-ELISA
Format 96T/48T
Assay time 2.0h
Detection range 1.56-100 ng/mL
Sensitivity 0.94 ng/mL
Sample type &Sample volume serum, plasma and other biological fluids; 50μL
Specificity This kit recognizes 3-NT in samples. No significant cross-reactivity or interference between 3-NT and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of 3-NT concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with 3-NT. During the reaction, 3-NT in the sample or standard competes with a fixed amount of 3-NT on the solid phase supporter for sites on the Biotinylated Detection Ab specific to 3-NT. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of 3-NT in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
ItemSpecificationsStorage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips
-20℃, 6 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 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 1 vial, 10 mL 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8℃
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level 3-NT were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level 3-NT 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 (ng/mL) 5.00 12.50 38.30 5.10 13.10 36.60
Standard deviation 0.30 0.70 1.80 0.30 0.60 1.90
CV (%) 6.00 5.60 4.70 5.88 4.58 5.19

Recovery

The recovery of 3-NT 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=8) 97-108 102
EDTA plasma(n=8) 91-108 99
Cell culture media(n=8) 93-105 100

Linearity

Samples were spiked with high concentrations of 3-NT and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Synonyms3-Nitro-L-Tyrosine, Nitrotyrosine

Assay Procedures

elisa assay procedure 1

1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C

elisa assay procedure 2

2. Aspirate and wash the plate for 3 times

elisa assay procedure 3

3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 4

4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 5

5. Add 50μL Stop Solution

elisa assay procedure 6

6. Read the plate at 450nm immediately. Calculation of the results

Related Products

Elabscience® provides ancillary reagents required for ELISA kit developments.11 reagents in total, including reagents for coating, blocking, washing, improving the stability and reducing matrix interference etc.
Cat.No. Product Name Specifications
E-ELIR-001 ELISA Plate Coating Buffer(1×) 100mL, 1000mL
E-ELIR-002 ELISA Plate Coating Buffer(5×) 100mL, 1000mL
E-ELIR-003 ELISA Plate Blocking Buffer 100mL, 1000mL
E-ELIR-004 Wash Buffer for Sandwich-ELISA(25×) 100mL, 1000mL
E-ELIR-005 Wash Buffer for Competitive-ELISA(25×) 100mL, 1000mL
E-ELIR-006 Stop Solution 100mL, 1000mL
E-ELIR-007 HRP-conjugate Stabilizer 10mL, 100mL, 500mL
E-ELIR-008 HRP-conjugate Diluent 100mL, 1000mL
E-ELIR-009 Biotinylated Antibody Stabilizer 10mL, 100mL, 500mL
E-ELIR-0010 Biotinylated Antibody Diluent 10mL, 100mL, 500mL
E-ELIR-0011 Sample Diluent 10mL, 100mL, 500mL

Citations

From now on, if you have published a paper by using any of our products since 1/1/2019, fill out the “Elabscience Publication Reward Application Form”carefully and send it to orders@elabscience.com, we will get back to you with the reward after we confirm it ASAP!

View more details about our Publication Reward >>

  1. Oxidative Medicine and Cellular Longevity (2020) IF: 5.076
    Vagus Nerve Stimulation Ameliorates Renal Ischemia-Reperfusion Injury through Inhibiting NF- κ B Activation and iNOS Protein Expression

    DOI: 10.1155/2020/7106525

    PMID: 32148655

    Sample: Tissue homogenate
  2. Archives of Biochemistry and Biophysics (2019) IF: 3.118
    Effect of a high sucrose and high fat diet in BDNF (+/-) mice on oxidative stress markers in adipose tissues

    DOI: 10.1016/j.abb.2019.02.004

    PMID: 30797748

    Sample: Serum
  3. Andrology (2020) IF: 2.86
    Low androgen status inhibits erectile function by inducing eNOS uncoupling in rat corpus cavernosum

    DOI: 10.1111/andr.12844

    PMID: 32594586

    Sample: Tissue homogenate
  4. Molecular vision (2018) IF: 2.219
    Protective effect of magnesium acetyltaurate and taurine against NMDA-induced retinal damage involves reduced nitrosative stress

    DOI: 99999999

    PMID: 30090013

    Sample: Tissue homogenate
  5. Biological trace element research (2018) IF: 2.361
    Melatonin Ameliorates Neuropharmacological and Neurobiochemical Alterations Induced by Subchronic Exposure to Arsenic in Wistar Rats

    DOI: 10.1007/s12011-018-1537-1

    PMID: 30306420

    Sample: Tissue homogenate
  6. Plos One (2017) IF: 2.806
    Reduction Of Oxidative-Nitrosative Stress Underlies Anticataract Effect Of Topically Applied Tocotrienol In Streptozotocin-Induced Diabetic Rats.

    DOI: 10.1371/journal.pone.0174542

    PMID: 28350848

    Sample: Tissue homogenate
  7. International Journal of Mycobacteriology (2020)
    Plasma levels of tumor necrosis factor-alpha, interferon-gamma, inducible nitric oxide synthase, and 3-nitrotyrosine in drug-resistant and drug-sensitive pulmonary tuberculosis patients, Ibadan, Nigeria

    DOI: 10.4103/ijmy.ijmy_63_20

    PMID: 32474541

    Sample: Plasma
  8. Nigerian journal of clinical practice (2019) IF: 0.713
    Markers of lipid and protein peroxidation among Nigerian university students with dysmenorrhea

    DOI: 10.4103/njcp.njcp_279_18

    PMID: 30729939

    Sample: Plasma
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