Multiplex Assay Kit for Thyrotropin Releasing Hormone (TRH) ,etc. by FLIA (Flow Luminescence Immunoassay)

TRF; Thyrotropin-Releasing Factor; Thyroliberin; Protirelin; Pro-tirelin; Prothyroliberin; TSH-releasing factor

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Thyrotropin Releasing Hormone (TRH) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Thyrotropin Releasing Hormone (TRH) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Thyrotropin Releasing Hormone (TRH) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Thyrotropin Releasing Hormone (TRH) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 88-102 93
EDTA plasma(n=5) 92-101 97
heparin plasma(n=5) 85-95 89
sodium citrate plasma(n=5) 89-99 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Thyrotropin Releasing Hormone (TRH) ,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 Thyrotropin Releasing Hormone (TRH) ,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 Thyrotropin Releasing Hormone (TRH) ,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) 78-98% 95-102% 91-101% 93-101%
EDTA plasma(n=5) 96-103% 93-103% 86-94% 78-103%
heparin plasma(n=5) 82-90% 80-99% 95-105% 86-98%
sodium citrate plasma(n=5) 83-94% 79-102% 87-99% 83-101%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:TRH) 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 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Plos one Thyrotropin-Releasing Hormone (TRH) Promotes Wound Re-Epithelialisation in Frog and Human Skin PubMed: PMC3759422
Analyst Ultrasensitive detection of thyrotropin-releasing hormone based on azo coupling and surface-enhanced resonance Raman spectroscopy Pubmed:27338554
journal of neuroendocrinology Thyroid dysfunction in children with autism spectrum disorder is associated with folate receptor α autoimmune disorder. pubmed:28199771
Journal of Neuroendocrinology Thyroid dysfunction in children with autism spectrum disorder is associated with folate receptor α autoimmune disorder 10.1111/jne.12461
Frontiers in Molecular Neuroscience Impact of Triclosan on Female Reproduction through Reducing Thyroid Hormones to Suppress Hypothalamic Kisspeptin Neurons in Mice Pubmed:29403355
Cell Reports Zika Virus Infection in Hypothalamus Causes Hormone Deficiencies and Leads to Irreversible Growth Delay and Memory Impairment in Mice Pubmed: 30404008
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