Instant ELISA Kit for Isocitrate Dehydrogenase 1, Soluble (IDH1)

PICD; IDP; Oxalosuccinate decarboxylase; NADP(+)-specific ICDH; Cytosolic NADP-isocitrate dehydrogenase

Specificity

This assay has high sensitivity and excellent specificity for detection of Instant Isocitrate Dehydrogenase 1, Soluble (IDH1).
No significant cross-reactivity or interference between Instant Isocitrate Dehydrogenase 1, Soluble (IDH1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Instant Isocitrate Dehydrogenase 1, Soluble (IDH1) and the recovery rates were calculated by comparing the measured value to the expected amount of Instant Isocitrate Dehydrogenase 1, Soluble (IDH1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 87-101 97
EDTA plasma(n=5) 96-103 101
heparin plasma(n=5) 80-102 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Instant Isocitrate Dehydrogenase 1, Soluble (IDH1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Instant Isocitrate Dehydrogenase 1, Soluble (IDH1) 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 Instant Isocitrate Dehydrogenase 1, Soluble (IDH1) 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) 82-102% 90-97% 84-97% 90-103%
EDTA plasma(n=5) 89-98% 78-104% 79-91% 87-94%
heparin plasma(n=5) 90-97% 83-95% 88-95% 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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 5 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 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 30 minutes at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 30 minutes at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 10 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
Molecular & Cellular Proteomics Identification of IDH1 as a Potential Diagnostic and Prognostic Biomarker for Non-small-cell Lung Cancer by Proteomic Analysis Mcponline: 008821
Molecular & Cellular Proteomics Identification of Isocitrate Dehydrogenase 1 as a Potential Diagnostic and Prognostic Biomarker for Non-small Cell Lung Cancer by Proteomic Analysis Pubmed: 22064513
Clinical Cancer Research Isocitrate dehydrogenase 1 is a novel plasma biomarker for the diagnosis of non-small cell lung cancer Pubmed: 24046070
Onco Targets Ther The effects of PGC-1α on the proliferation and energy metabolism of malignant endometrial cancer cells PubMed: 25914546
Oncogene Metformin sensitizes endometrial cancer cells to chemotherapy through IDH1-induced Nrf2 expression via an epigenetic mechanism Pubmed:29921847
Molecular Oncology IDH 2 is a novel diagnostic and prognostic serum biomarker for non‐small‐cell lung cancer Pubmed:29465809
Cancers Serological Biomarkers for Early Detection of Hepatocellular Carcinoma: A Focus on Autoantibodies against Tumor-Associated Antigens Encoded by Cancer Driver … Pubmed: 32443439
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