Multiplex Assay Kit for Netrin 1 (Ntn1) ,etc. by FLIA (Flow Luminescence Immunoassay)

NTN1L; Epididymis tissue protein Li 131P

(Note: Up to 8-plex in one testing reaction)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 79-104 83
EDTA plasma(n=5) 91-101 96
heparin plasma(n=5) 92-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Netrin 1 (Ntn1) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 93-101% 87-102% 97-104% 84-95%
EDTA plasma(n=5) 89-97% 81-89% 89-99% 86-93%
heparin plasma(n=5) 80-99% 89-103% 79-94% 79-91%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:Ntn1) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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 Chicken (Gallus) Organism species Applications (RESEARCH USE ONLY!)
RPB827Ga01 Recombinant Netrin 1 (Ntn1) Positive Control; Immunogen; SDS-PAGE; WB.
PAB827Ga01 Polyclonal Antibody to Netrin 1 (Ntn1) WB; IHC; ICC; IP.
MAB827Ga21 Monoclonal Antibody to Netrin 1 (Ntn1) WB; IHC; ICC; IP.
SEB827Ga ELISA Kit for Netrin 1 (Ntn1) Enzyme-linked immunosorbent assay for Antigen Detection.
LMB827Ga Multiplex Assay Kit for Netrin 1 (Ntn1) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.