ELISA Kit for NADPH Oxidase 4 (NOX4)

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

Specificity

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

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level NADPH Oxidase 4 (NOX4) 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) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

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
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
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