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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) 80-94 83
EDTA plasma(n=5) 91-102 97
heparin plasma(n=5) 81-99 87

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) 89-105% 99-105% 96-105% 98-105%
EDTA plasma(n=5) 93-101% 88-105% 96-105% 78-92%
heparin plasma(n=5) 92-104% 89-97% 79-101% 88-104%

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 Reference
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
NATURE MEDICINE 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
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.