ELISA Kit for Netrin 1 (Ntn1)

NTN1L; Epididymis tissue protein Li 131P

Specificity

This assay has high sensitivity and excellent specificity for detection of Netrin 1 (Ntn1).
No significant cross-reactivity or interference between Netrin 1 (Ntn1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Netrin 1 (Ntn1) and the recovery rates were calculated by comparing the measured value to the expected amount of Netrin 1 (Ntn1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-98 92
EDTA plasma(n=5) 96-103 101
heparin plasma(n=5) 79-101 83

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Netrin 1 (Ntn1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Netrin 1 (Ntn1) 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 Netrin 1 (Ntn1) 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) 99-105% 78-103% 82-91% 87-94%
EDTA plasma(n=5) 85-101% 78-104% 78-102% 88-101%
heparin plasma(n=5) 94-101% 86-103% 87-101% 80-98%

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 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

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

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Magazine Citations
Transplantation Proceedings Netrin-1: A Novel Universal Biomarker of Human Kidney Injury PubMed: 20620466
Biomarkers Plasma netrin-1 is a diagnostic biomarker of human cancers PubMed: PMC3143477
Journal of Maternal-Fetal and Neonatal Medicine Cord blood netrin-1 and -4 concentrations in term pregnancies with normal, restricted and increased fetal growth Informahealthcare: Source
4 Netrin-1 promotes adipose tissue macrophage retention and insulin resistance in obesity Pubmed:24584118
J Matern?Fetal?Neonatal Med.? Cord blood netrin-1 and-4 concentrations in term pregnancies with normal, restricted and increased fetal growth Pubmed:24716747
Renal failure. Urinary netrin-1 and KIM-1 as early biomarkers for septic acute kidney injury Pubmed:25154466
Stem Cell Research & Therapy Netrin-1 acts as a non-canonical angiogenic factor produced by human Wharton’s jelly mesenchymal stem cells (WJ-MSC) articles:10.1186
Oncotarget The effect of aspirin on circulating netrin-1 levels in humans is dependent on the inflammatory status of the vascular endothelium. pubmed:29156815
Frontiers in Cellular Neuroscience Neuroprotection Exerted by Netrin-1 and Kinesin Motor KIF1A in Secondary Brain Injury following Experimental Intracerebral Hemorrhage in Rats Pubmed:29375318
Scientific Reports Netrin-1 promotes diabetic corneal wound healing through molecular mechanisms mediated via the adenosine 2B receptor Pubmed:29662125
Family Practice and Palliative Care Relationship of serum netrin-1 levels with breast masses Doi: 10.22391/fppc.467700
JOURNAL OF BIOLOGICAL CHEMISTRY Role of microRNA in CB1 antagonist-mediated regulation of adipose tissue macrophage polarization and chemotaxis during diet-induced obesity Pubmed: 30910812
International Journal of Molecular Sciences Downregulation of the Netrin-1 Receptor UNC5b Underlies Increased Placental Angiogenesis in Human Gestational Diabetes Mellitus Pubmed: 30897795
Cancer Biomark Serum netrin-1 as a biomarker for colorectal cancer detection Pubmed: 32474463
Níveis Elevados de Netrina-1 e IL-1β em Mulheres Idosas com SCA: Pior Prognóstico no Acompanhamento de Dois Anos
Tuberk Toraks Association of plasma netrin-1 level with inflammatory balance and comorbidities in patients with acute exacerbation of COPD 33853303
Adv Healthc Mater Conductive Hydrogel Conduits with Growth Factor Gradients for Peripheral Nerve Repair in Diabetics with Non‐suture Tape Pubmed:35670309
Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
RPB827Ra01 Recombinant Netrin 1 (Ntn1) Positive Control; Immunogen; SDS-PAGE; WB.
PAB827Ra01 Polyclonal Antibody to Netrin 1 (Ntn1) WB; IHC; ICC; IP.
MAB827Ra21 Monoclonal Antibody to Netrin 1 (Ntn1) WB; IHC; ICC; IP.
SEB827Ra ELISA Kit for Netrin 1 (Ntn1) Enzyme-linked immunosorbent assay for Antigen Detection.
LMB827Ra Multiplex Assay Kit for Netrin 1 (Ntn1) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.