Multiplex Assay Kit for NADPH Oxidase 4 (NOX4) ,etc. by FLIA (Flow Luminescence Immunoassay)

KOX; KOX-1; RENOX; Nicotinamide Adenine Dinucleotide Phosphate Oxidase 4

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

Specificity

This assay has high sensitivity and excellent specificity for detection of NADPH Oxidase 4 (NOX4) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between NADPH Oxidase 4 (NOX4) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant NADPH Oxidase 4 (NOX4) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of NADPH Oxidase 4 (NOX4) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 96-103 99
EDTA plasma(n=5) 81-93 87
heparin plasma(n=5) 98-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level NADPH Oxidase 4 (NOX4) ,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 NADPH Oxidase 4 (NOX4) ,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 NADPH Oxidase 4 (NOX4) ,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) 84-102% 98-105% 85-95% 83-94%
EDTA plasma(n=5) 95-103% 89-102% 83-101% 84-101%
heparin plasma(n=5) 80-88% 90-104% 98-105% 93-101%

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#:NOX4) 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
Chem Biol Interact Apocynin protects against ethanol-induced gastric ulcer in rats by attenuating the upregulation of NADPH oxidases 1 and 4 PubMed: 26522475
Prostate Oxidative stress promotes benign prostatic hyperplasia PubMed: 26417670
PLoS One. Effects of Single and Combined Losartan and Tempol Treatments on Oxidative Stress, Kidney Structure and Function in Spontaneously Hypertensive Rats with Early Course of Proteinuric Nephropathy. pubmed:27560781
JOURNAL OF THE AMERICAN SOCIETY OF HYPERTENSION Galectin-3 mediates the pulmonary arterial hypertension-induced right ventricular remodeling through interacting with NADPH oxidase 4. pubmed:28431936
Experimental and Therapeutic Medicine Chronic restraint stress induces esophageal fibrosis with enhanced oxidative stress in a murine model
PLoS Biol NOX5-induced uncoupling of endothelial NO synthase is a causal mechanism and theragnostic target of an age-related hypertension endotype Pubmed: 33170835
Free Radic Biol Med Biphasic changes in TGF-β1 signaling drive NSAID-induced multi-organ damage Pubmed: 32750407
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
RPB924Hu02 Recombinant NADPH Oxidase 4 (NOX4) Positive Control; Immunogen; SDS-PAGE; WB.
RPB924Hu01 Recombinant NADPH Oxidase 4 (NOX4) Positive Control; Immunogen; SDS-PAGE; WB.
PAB924Hu01 Polyclonal Antibody to NADPH Oxidase 4 (NOX4) WB; IHC; ICC; IP.
PAB924Hu02 Polyclonal Antibody to NADPH Oxidase 4 (NOX4) WB; IHC; ICC; IP.
LAB924Hu71 Biotin-Linked Polyclonal Antibody to NADPH Oxidase 4 (NOX4) WB; IHC; ICC.
MAB924Hu22 Monoclonal Antibody to NADPH Oxidase 4 (NOX4) WB; IHC; ICC; IP.
SEB924Hu ELISA Kit for NADPH Oxidase 4 (NOX4) Enzyme-linked immunosorbent assay for Antigen Detection.
MEB924Hu Mini Samples ELISA Kit for NADPH Oxidase 4 (NOX4) Enzyme-linked immunosorbent assay for Antigen Detection.
LMB924Hu Multiplex Assay Kit for NADPH Oxidase 4 (NOX4) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.