Multiplex Assay Kit for Isocitrate Dehydrogenase 1, Soluble (IDH1) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

(Note: Up to 8-plex in one testing reaction)

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 85-99 89
EDTA plasma(n=5) 79-91 84
heparin plasma(n=5) 86-101 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Isocitrate Dehydrogenase 1, Soluble (IDH1) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 FLIA (Flow Luminescence Immunoassay) 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 FLIA (Flow Luminescence Immunoassay) 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) 95-103% 98-105% 89-98% 82-102%
EDTA plasma(n=5) 79-101% 85-101% 78-104% 95-105%
heparin plasma(n=5) 85-97% 83-99% 78-95% 87-94%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:IDH1) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 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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RPH839Ra01 Recombinant Isocitrate Dehydrogenase 1, Soluble (IDH1) Positive Control; Immunogen; SDS-PAGE; WB.
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MAH839Ra21 Monoclonal Antibody to Isocitrate Dehydrogenase 1, Soluble (IDH1) WB; IHC; ICC; IP.
SEH839Ra ELISA Kit for Isocitrate Dehydrogenase 1, Soluble (IDH1) Enzyme-linked immunosorbent assay for Antigen Detection.
LMH839Ra Multiplex Assay Kit for Isocitrate Dehydrogenase 1, Soluble (IDH1) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.