Multiplex Assay Kit for Lysyl Oxidase (LOX) ,etc. by FLIA (Flow Luminescence Immunoassay)

Protein-Lysine 6-Oxidase

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 93-105 101
EDTA plasma(n=5) 84-104 99
heparin plasma(n=5) 96-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Lysyl Oxidase (LOX) ,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 Lysyl Oxidase (LOX) ,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 Lysyl Oxidase (LOX) ,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) 87-96% 89-103% 80-89% 89-104%
EDTA plasma(n=5) 90-101% 94-104% 93-102% 84-101%
heparin plasma(n=5) 91-104% 97-104% 93-102% 98-105%

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#:LOX) 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
American Journal of Hematology The expression of lysyl-oxidase gene family members in myeloproliferative neoplasms. PubMed: 23494965
Current Eye Research High Glucose Induced Differential Expression of Lysyl Oxidase and Its Isoform in ARPE-19 Cells Pubmed: 22966818
European Journal of Surgical Oncology Evaluation of serum lysyl oxidase as a blood test for colorectal cancer Pubmed: 24246612
Vaccine Copia autorizada por CDR Ingentaconnect: Source
Arthritis & Rheumatology Lysyl oxidase is a potential biomarker of fibrosis in systemic sclerosis Pubmed: 24574233
Eur J Surg Oncol.? Evaluation of serum lysyl oxidase as a blood test for colorectal cancer Pubmed:Pmc4237001
BMC Cancer. Ionizing radiation induces tumor cell lysyl oxidase secretion Pubmed:Pmc4223762
DNA Cell Biol. Lysyl Oxidase Genetic Variants Affect Gene Expression in Cervical Cancer Pubmed:24945327
scandinavian journal of immunology Increased Serum Levels of the IL-33 Neutralizing sST2 in Limited Cutaneous Systemic Sclerosis PubMed: 26095613
Curr Eye Res Correlation of Aqueous Humor Lysyl Oxidase Activity with TGF-ß Levels and LOXL1 Genotype in Pseudoexfoliation Pubmed:27116380
journal of bone and mineral metabolism LOX Fails to Substitute for RANKL in Osteoclastogenesis. pubmed:27606829
38 Blocking Surgically Induced Lysyl Oxidase Activity Reduces the Risk of Lung Metastases. pubmed:28445728
Molecular Medicine Reports Lysyl oxidase is involved in synovial hyperplasia and angiogenesis in rats with collagen‑induced arthritis pubmed:28901438
Revista Española de Cardiología (English Edition) A Global Assessment of Circulating Prolysyl Oxidase in Nonischemic Patients With Garden-variety Heart Failure With Preserved Ejection Fraction Pubmed:29807761
Molecular Medicine Reports Lysyl oxidase inhibition via β-aminoproprionitrile hampers human umbilical vein endothelial cell angiogenesis and migration in vitro Pubmed:29393489
Thyroid Lysyl Oxidase is a key player in BRAF/MAPK pathway-driven thyroid cancer aggressiveness Pubmed: 30398411
Am J Physiol Lung Cell Mol Physiol Lysyl Oxidase Directly Contributes to Extracellular Matrix Production and Fibrosis in Systemic Sclerosis. Pubmed: 33026236
J Am Heart Assoc Collagen Cross©\Linking Is Associated With Cardiac Remodeling in Hypertrophic Obstructive Cardiomyopathy 33356379
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