ELISA Kit for Sorbitol Dehydrogenase (SDH)

SORD; L-iditol 2-dehydrogenase

Specificity

This assay has high sensitivity and excellent specificity for detection of Sorbitol Dehydrogenase (SDH).
No significant cross-reactivity or interference between Sorbitol Dehydrogenase (SDH) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Sorbitol Dehydrogenase (SDH) and the recovery rates were calculated by comparing the measured value to the expected amount of Sorbitol Dehydrogenase (SDH) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 91-101 96
EDTA plasma(n=5) 78-101 92
heparin plasma(n=5) 93-101 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Sorbitol Dehydrogenase (SDH) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Sorbitol Dehydrogenase (SDH) 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 Sorbitol Dehydrogenase (SDH) 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) 78-101% 93-101% 88-95% 92-105%
EDTA plasma(n=5) 82-96% 84-92% 95-102% 79-92%
heparin plasma(n=5) 78-94% 88-96% 98-105% 78-92%

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
Kafkas Univ Vet Fak Derg The Importance of Concentrations of Sorbitol Dehydrogenase and Glutamate Dehydrogenase and B-Mode Ultrasonographic Examination in The Diagnosis of Hepatic Lipidosis in Dairy Cows Kafkas: Source
Journal of Food Science and Technology Grape seed extract and Zinc containing nutritional food supplement delays onset and progression of Streptozocin-induced diabetic cataract in Wistar rats Springer: Source
Clinical and Experimental Optometry Aldo-keto reductase and sorbitol dehydrogenase enzymes in Egyptian diabetic patients with and without proliferative diabetic retinopathy. Pubmed: 23452182
Biomedicine & Pharmacotherapy Zinc/alogliptin combination attenuates testicular toxicity induced by doxorubicin in rats: Role of oxidative stress, apoptosis and TGF-β1/NF-κB signaling pubmed:29091894
Molecular Medicine Reports Protective effects of N (2)‑L‑alanyl‑L‑glutamine mediated by the JAK2/STAT3 signaling pathway on myocardial ischemia reperfusion Pubmed:29393473
Effect of the combination between empagliflozin and calcipotriol on cadmium-induced testicular toxicity in rats
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