ELISA Kit for Nesfatin 1 (NES1)

Specificity

This assay has high sensitivity and excellent specificity for detection of Nesfatin 1 (NES1).
No significant cross-reactivity or interference between Nesfatin 1 (NES1) and analogues was observed.

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) 80-101 86
EDTA plasma(n=5) 96-104 101
heparin plasma(n=5) 87-96 90

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) 90-98% 80-90% 96-103% 83-101%
EDTA plasma(n=5) 83-90% 96-103% 97-105% 84-103%
heparin plasma(n=5) 78-94% 82-89% 80-101% 87-96%

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.

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Magazine Citations
Bulletin of Clinical Psychopharmacology Plasma Nesfatin-1 Level May Be Associated with Disease Severity in Patients with Panic Disorder Psikofarmakoloji: 2043
Progress in Neuro-Psychopharmacology & Biological Psychiatry High plasma nesfatin-1 level in patients with major depressive disorder PubMed: 21163317
Endocrine Cord blood nesfatin-1 and apelin-36 levels in gestational diabetes mellitus SpringerLink: h978088657555w16
Bulletin of Clinical Psychopharmacology Increased Plasma Nesfatin-1 Levels in Patients with Obsessive Compulsive Disorder Psikofarmakoloji: 2212
Psychoneuroendocrinology. Decreased plasma nesfatin-1 levels in patients with generalized anxiety disorder PubMed: 22595767
Iranian Journal of Health and Physical activity Effects of Aerobic Training, with or without Zizyphus Jujuba Water Extraction, on Fundus Nesfatin-1, ATP, HDL-C, and LDL-C Concentrations in Female Rats Ijvst: Source
European Journal of Obstetrics & Gynecology and Reproductive Biology Decreased serum nesfatin-1 levels in endometriosis Pubmed: 24702902
Bulletin of Clinical Psychopharmacology Increased Plasma Nesfatin-1 Levels in Patients with Obsessive Compulsie Disorder. Psikofarmakoloji: Source
Current Pharmaceutical Design Role of NUCB2/nesfatin-1 as a possible biomarker. Pubmed: 23537082
Regulatory, Integrative and Comparative Physiology A role of nesfatin-1/NucB2 in dehydration-induced anorexia Pubmed:24829503
J Endocrinol Invest. Plasma nesfatin-1 levels are increased in patients with polycystic ovary syndrome Pubmed:24920281
Int J Surg. 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
Hormone molecular biology and clinical investigation The effects of short-term high-intensity interval training vs. moderate-intensity continuous training on plasma levels of nesfatin-1 and inflammatory markers Pubmed:25581765
Turk J Gastroenterol. New hormones to predict the severity of gallstone-induced acute pancreatitis Pubmed:25599787
Respiration Is There a Relationship between Obstructive Sleep Apnea Syndrome Severity and Nesfatin-1? PubMed: 26088934
J Clin Res Pediatr Endocninol Correlation of Brain Neuropeptide (Nesfatin-1 and Orexin-A) Concentrations with Anthropometric and Biochemical Parameters in Malnourished Children PubMed: 26831553
Bosn J Basic Med Sci Nesfatin-1 and Vitamin D levels may be associated with systolic and diastolic blood pressure values and hearth rate in polycystic ovary syndrome PubMed: 26295295
Original Article / Orijinal Makale Relationship and Interaction between Anaerobic Sports Branches and Serum Nesfatin-1 Sayilar: 294
Gynecol Endocrinol Nesfatin-1 levels and metabolic markers in polycystic ovary syndrome PubMed: 26062107
International Journal of Neuroscience Serum nesfatin-1 levels: a potential new biomarker in patients with subarachnoid hemorrhage* Pubmed:26872406
未找到 Comparing the nesfatin-1 levels between individuals with subjective cognitive decline and mild cognitive impairment P.1.e.012.pdf
International Journal of Developmental Serum nesfatin-1 levels: a potential new biomarker in patients with subarachnoid hemorrhage. pubmed:26872406
Current Pharmaceutical Design Metabolic and Cardiovascular Actions of Nesfatin-1: Implications in Health and Disease pubmed:28137218
Arch Endocrinol Metab Serum nesfatin-1 levels are decreased in pregnant women newly diagnosed with gestational diabetes pubmed:28977160
Indian Journal of Psychiatry Serum NUCB2/nesfatin-1 levels in different stages of alcohol dependence: Is there a relationship with craving? pubmed:28529367
Biomedicine & Pharmacotherapy Nesfatin-1 promotes VSMC migration and neointimal hyperplasia by upregulating matrix metalloproteinases and downregulating PPARγ Pubmed:29604590
Biochimica et Biophysica Acta Nesfatin-1 functions as a switch for phenotype transformation and proliferation of VSMCs in hypertensive vascular remodeling Pubmed:29627363
BMC Psychiatry Plasma nesfatin-1 level is associated with severity of depression in Chinese depressive patients Pubmed:29615007
Probiotics and Antimicrobial Proteins Mitra Zarrati, Mahsa Raji Lahiji, Eisa Salehi, Bahareh Yazdani, Elham Razmpoosh, Raheleh Shokouhi Shoormasti & Farzad Shidfar Pubmed: 30232744
An elevated serum level of nesfatin-1 may predict poor outcomes in patients with aneurysmal subarachnoid hemorrhages
clinical neurology and neurosurgery Relationshıp of spontaneous subarachnoid haemorrhage and cerebral aneurysm to serum Visfatin and Nesfatin-1 levels Pubmed: 32311618
Ann Nutr Metab Copeptin and Nesfatin-1 Are Interrelated Biomarkers with Roles in the Pathogenesis of Insulin Resistance in Chinese Children with Obesity Pubmed: 33027789
Endokrynologia Polska Assessing the blood concentration of new adipocytokines in patients with ischaemic stroke 33283259
Increased serum nesfatin-1 levels among adolescents diagnosed with major depressive disorder
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