ELISA Kit for Autophagy Related Protein 5 (ATG5)

ASP; APG5; APG5L; hAPG5; Apoptosis-specific protein; Autophagy protein 5

Specificity

This assay has high sensitivity and excellent specificity for detection of Autophagy Related Protein 5 (ATG5).
No significant cross-reactivity or interference between Autophagy Related Protein 5 (ATG5) and analogues was observed.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Autophagy Related Protein 5 (ATG5) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Autophagy Related Protein 5 (ATG5) 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
Frontiers in Cellular Neuroscience A Variant of the Autophagy-Related 5 Gene Is Associated with Child Cerebral Palsy. pubmed:29326554
Medical Science Monitor Relationship of Autophagy and Apoptosis with Total Occlusion of Coronary Arteries Pubmed: 30273932
Scientific Reports Plasma ATG5 is increased in Alzheimer's disease
Acta Neurol Belg Association of ATG5 gene polymorphism with Parkinson's disease in a Han Chinese population 34661876
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