Multiplex Assay Kit for Hypoxia Inducible Factor 2 Alpha (HIF2a) ,etc. by CBA (Cytometric Bead Array)

EPAS1; HLF; MOP2; PASD2; BHLHE73; Endothelial PAS Domain 1; HIF-1 Alpha-Like Factor; Basic-helix-loop-helix-PAS protein MOP2; Class E basic helix-loop-helix protein 73

Specificity

This assay has high sensitivity and excellent specificity for detection of Hypoxia Inducible Factor 2 Alpha (HIF2a) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Hypoxia Inducible Factor 2 Alpha (HIF2a) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Hypoxia Inducible Factor 2 Alpha (HIF2a) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Hypoxia Inducible Factor 2 Alpha (HIF2a) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-103 98
EDTA plasma(n=5) 83-101 92
heparin plasma(n=5) 80-96 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hypoxia Inducible Factor 2 Alpha (HIF2a) ,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 Hypoxia Inducible Factor 2 Alpha (HIF2a) ,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 Hypoxia Inducible Factor 2 Alpha (HIF2a) ,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) 98-105% 88-95% 81-105% 82-97%
EDTA plasma(n=5) 96-104% 78-99% 86-101% 79-104%
heparin plasma(n=5) 78-102% 81-89% 95-103% 92-101%

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

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
Neuro Oncol Combination viroimmunotherapy with checkpoint inhibition to treat glioma, based on location-specific tumor profiling PubMed: 26409567
Clinical Trials in Degenerative Diseases Correlating oxidative stress-related factors with bone metabolic markers in older adult male patients exhibiting degenerative osteoporosis in the high-altitude hypoxic area of China: study protocol for a non-randomized controlled trial issn:2542-3975
Journal of Physiological Anthropology Telomere elongation protects heart and lung tissue cells from fatal damage in rats exposed to severe hypoxia Pubmed:29454386
Oxidative damage and iron dyshomeostasis in thalassemic patients
Neuropsychiatric Disease and Treatment The Association Between Hypoxia Improvement and Electroconvulsive Therapy for Major Depressive Disorder 34588778
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