ELISA Kit for Nesfatin 1 (NES1) Rattus norvegicus (Rat) Competition ELISA

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  • ELISA Kit for Nesfatin 1 (NES1) Packages (Simulation)
  • ELISA Kit for Nesfatin 1 (NES1) Packages (Simulation)
  • ELISA Kit for Nesfatin 1 (NES1) Results demonstration
  • CEA242Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Nesfatin 1 (NES1) and the recovery rates were calculated by comparing the measured value to the expected amount of Nesfatin 1 (NES1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 88-96 92
EDTA plasma(n=5) 78-104 83
heparin plasma(n=5) 91-105 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Nesfatin 1 (NES1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Nesfatin 1 (NES1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Nesfatin 1 (NES1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 92-99% 99-105% 88-95% 86-94%
EDTA plasma(n=5) 83-90% 79-89% 82-96% 93-103%
heparin plasma(n=5) 86-94% 80-96% 87-103% 93-101%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

ELISA Kit for Nesfatin 1 (NES1)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Nesfatin 1 (NES1) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Nesfatin 1 (NES1) and unlabeled Nesfatin 1 (NES1) (Standards or samples) with the pre-coated antibody specific to Nesfatin 1 (NES1). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Nesfatin 1 (NES1) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Nesfatin 1 (NES1) in the sample.

Citations

  • Plasma Nesfatin-1 Level May Be Associated with Disease Severity in Patients with Panic DisorderPsikofarmakoloji: 2043
  • High plasma nesfatin-1 level in patients with major depressive disorderPubMed: 21163317
  • Cord blood nesfatin-1 and apelin-36 levels in gestational diabetes mellitusSpringerLink: h978088657555w16
  • Increased Plasma Nesfatin-1 Levels in Patients with Obsessive Compulsive Disorder Psikofarmakoloji: 2212
  • Decreased plasma nesfatin-1 levels in patients with generalized anxiety disorderPubMed: 22595767
  • Effects of Aerobic Training, with or without Zizyphus Jujuba Water Extraction, on Fundus Nesfatin-1, ATP, HDL-C, and LDL-C Concentrations in Female RatsIjvst: Source
  • Decreased serum nesfatin-1 levels in endometriosisPubmed: 24702902
  • Increased Plasma Nesfatin-1 Levels in Patients with Obsessive Compulsie Disorder.Psikofarmakoloji: Source
  • Role of NUCB2/nesfatin-1 as a possible biomarker.Pubmed: 23537082
  • A role of nesfatin-1/NucB2 in dehydration-induced anorexiaPubmed:24829503
  • Plasma nesfatin-1 levels are increased in patients with polycystic ovary syndromePubmed:24920281
  • The potential role of BMI, plasma leptin, nesfatin-1 and ghrelin levels in the early detection of pancreatic necrosis and severe acute pancreatitis: a prospective cohort study.Pubmed:25448651
  • The effects of short-term high-intensity interval training vs. moderate-intensity continuous training on plasma levels of nesfatin-1 and inflammatory markersPubmed:25581765
  • New hormones to predict the severity of gallstone-induced acute pancreatitisPubmed:25599787
  • Is There a Relationship between Obstructive Sleep Apnea Syndrome Severity and Nesfatin-1?PubMed: 26088934
  • Correlation of Brain Neuropeptide (Nesfatin-1 and Orexin-A) Concentrations with Anthropometric and Biochemical Parameters in Malnourished ChildrenPubMed: 26831553
  • Nesfatin-1 and Vitamin D levels may be associated with systolic and diastolic blood pressure values and hearth rate in polycystic ovary syndromePubMed: 26295295
  • Relationship and Interaction between Anaerobic Sports Branches and Serum Nesfatin-1Sayilar: 294
  • Nesfatin-1 levels and metabolic markers in polycystic ovary syndromePubMed: 26062107
  • Serum nesfatin-1 levels: a potential new biomarker in patients with subarachnoid hemorrhage*Pubmed:26872406
  • Comparing the nesfatin-1 levels between individuals with subjective cognitive decline and mild cognitive impairmentP.1.e.012.pdf
  • Serum nesfatin-1 levels: a potential new biomarker in patients with subarachnoid hemorrhage.pubmed:26872406
  • Metabolic and Cardiovascular Actions of Nesfatin-1: Implications in Health and Diseasepubmed:28137218
  • Serum nesfatin-1 levels are decreased in pregnant women newly diagnosed with gestational diabetespubmed:28977160
  • Serum NUCB2/nesfatin-1 levels in different stages of alcohol dependence: Is there a relationship with craving?pubmed:28529367
  • Nesfatin-1 promotes VSMC migration and neointimal hyperplasia by upregulating matrix metalloproteinases and downregulating PPARγPubmed:29604590
  • Nesfatin-1 functions as a switch for phenotype transformation and proliferation of VSMCs in hypertensive vascular remodelingPubmed:29627363
  • Plasma nesfatin-1 level is associated with severity of depression in Chinese depressive patientsPubmed:29615007
  • Mitra Zarrati, Mahsa Raji Lahiji, Eisa Salehi, Bahareh Yazdani, Elham Razmpoosh, Raheleh Shokouhi Shoormasti & Farzad ShidfarPubmed: 30232744
  • An elevated serum level of nesfatin-1 may predict poor outcomes in patients with aneurysmal subarachnoid hemorrhages
  • Relationshıp of spontaneous subarachnoid haemorrhage and cerebral aneurysm to serum Visfatin and Nesfatin-1 levelsPubmed: 32311618
  • Copeptin and Nesfatin-1 Are Interrelated Biomarkers with Roles in the Pathogenesis of Insulin Resistance in Chinese Children with ObesityPubmed: 33027789
  • Assessing the blood concentration of new adipocytokines in patients with ischaemic stroke33283259
  • Increased serum nesfatin-1 levels among adolescents diagnosed with major depressive disorder

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