Multiplex Assay Kit for Isocitrate Dehydrogenase 1, Soluble (IDH1) ,etc. by CBA (Cytometric Bead Array)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 102
EDTA plasma(n=5) 80-99 89
heparin plasma(n=5) 93-101 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Isocitrate Dehydrogenase 1, Soluble (IDH1) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Isocitrate Dehydrogenase 1, Soluble (IDH1) ,etc. by CBA (Cytometric Bead Array) 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 Isocitrate Dehydrogenase 1, Soluble (IDH1) ,etc. by CBA (Cytometric Bead Array) 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) 85-102% 87-101% 88-104% 87-94%
EDTA plasma(n=5) 92-105% 96-105% 86-101% 78-94%
heparin plasma(n=5) 92-102% 80-102% 85-99% 98-105%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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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
International Journal of Cancer Proteome profiling of enzalutamide‐resistant cell lines and serum analysis identified ALCAM as marker of resistance in castration‐resistant prostate cancer Pubmed:35689436
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