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

GRF; GHRF; Somatocrinin; Growth-Hormone-Releasing Factor; Sermorelin; Somatoliberin

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-104 87
EDTA plasma(n=5) 97-104 101
heparin plasma(n=5) 84-99 94
sodium citrate plasma(n=5) 78-102 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Growth Hormone Releasing Hormone (GHRH) ,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 Growth Hormone Releasing Hormone (GHRH) ,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 Growth Hormone Releasing Hormone (GHRH) ,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) 83-94% 86-94% 86-101% 98-105%
EDTA plasma(n=5) 97-105% 94-102% 97-104% 95-102%
heparin plasma(n=5) 78-102% 97-105% 78-90% 78-102%
sodium citrate plasma(n=5) 94-103% 90-102% 79-104% 97-105%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:GHRH) 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
19 Role of GHRH in Sleep and Growth Impairments Induced by Upper Airway Obstruction in Rats Ers: 00197610
19 Role of growth hormone-releasing hormone in sleep and growth impairments induced by upper airway obstruction in rats PubMed: 21406516
Global Veterinaria Effect of Moringa Oleifera Extract on Nicotine Induced Neurotoxicity in Female Rat and Their Embryos Idosi: Source
Cell Reports Zika Virus Infection in Hypothalamus Causes Hormone Deficiencies and Leads to Irreversible Growth Delay and Memory Impairment in Mice Pubmed: 30404008
Drug Testing and Analysis An Immuno‐PCR screen for the detection of CJC‐1295 and other Growth Hormone Releasing Hormone analogues in equine plasma Pubmed: 30489688
Sleep and Breathing Effects of somatotropic axis on cognitive dysfunction of obstructive sleep apnea Pubmed: 31073904
Diabetes Lack of brain insulin receptor substrate-1 causes growth retardation, with decreased expression of growth hormone-releasing hormone in the hypothalamus. 33980693
Sci Rep Associations between serum levels of brain-derived neurotrophic factor, corticotropin releasing hormone and mental distress in vitiligo patients Pubmed:35508633
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