Multiplex Assay Kit for Gonadotropin Releasing Hormone (GnRH) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

GNRH1; GRH; LNRH; LHRH; LH-RH I; Luliberin; Luteinizing-Hormone Releasing Hormone; Progonadoliberin-1; Gonadoliberin I; Gonadorelin; GnRH-associated peptide 1

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

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  • Multiplex Assay Kit for Gonadotropin Releasing Hormone (GnRH) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Gonadotropin Releasing Hormone (GnRH) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Gonadotropin Releasing Hormone (GnRH) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA843Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 86-104 101
EDTA plasma(n=5) 86-99 93
heparin plasma(n=5) 99-105 102
sodium citrate plasma(n=5) 83-93 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Gonadotropin Releasing Hormone (GnRH) ,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 Gonadotropin Releasing Hormone (GnRH) ,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 Gonadotropin Releasing Hormone (GnRH) ,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) 92-103% 79-93% 96-105% 98-105%
EDTA plasma(n=5) 84-92% 78-88% 85-96% 91-105%
heparin plasma(n=5) 88-104% 85-104% 86-103% 87-102%
sodium citrate plasma(n=5) 84-97% 94-102% 95-102% 80-88%

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#:GnRH) 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.

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

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards,Labeled antigen and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest.A competitive inhibition reaction is launched between biotin labeled analytes of interest and unlabeled analytes of interest (Standards or samples) with the pre-coated antibody specific to analytes of interest. Following a wash to remove any unbound substances, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE) is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer. The MFI developed is reverse proportional to the concentration of analytes of interest in the sample.

Giveaways

Citations

  • Maternal exposure to dioxin imprints sexual immaturity of the pups through fixing the status of the reduced expression of hypothalamic gonadotropin-releasing …Pubmed:24132183
  • Heavy Resistance Training and Supplementation With the Alleged Testosterone Booster Nmda has No Effect on Body Composition, Muscle Performance, and Serum Hormones Associated With the Hypothalamo-Pituitary-Gonadal Axis in Resistance-Trained MalesPubmed:24570624
  • Perfluorooctane sulfonate effects on the reproductive axis in adult male ratsPubmed:25171141
  • Possible role of serotonin and neuropeptide Y on the disruption of the reproductive axis activity by perfluorooctane sulfonatePubmed:25623392
  • Chronic exposure of female mice to an environmental level of perfluorooctane sulfonate suppresses estrogen synthesis through reduced histone H3K14 acetylation of the StAR promoter leading to deficits in follicular development and ovulationPubMed: 26358002
  • Overexpression of PRL7D1 in Leydig Cells Causes Male Reproductive Dysfunction in MicePubmed:26771609
  • OP0019-HPR Effectiveness of A Progressive Resistance Strength Program on Hand Osteoarthritis: A Randomised Controlled Trial60.1.abstract
  • OP0021-HPR Sleep Disorders in Patients with Rheumatoid Arthritis and Relationship with Disease Activity60.3.abstract
  • Exposure of pregnant mice to perfluorobutanesulfonate causes hypothyroxinemia and developmental abnormalities in female offspringpubmed:27803384
  • OP0020 The Association of Gonadotropin-Releasing Hormone and Cytokines in Rheumatoid Arthritiscontent:75
  • Pyrethroid Insecticide Cypermethrin Accelerates Pubertal Onset in Male Mice via Disrupting Hypothalamic-Pituitary-Gonadal Axispubmed:28731686
  • Impact of Perfluorooctane Sulfonate on Reproductive Ability of Female Mice through Suppression of Estrogen Receptor a-Activated Kisspeptin Neurons10.1093:toxsci
  • Nectin-like molecule 2, a necessary sexual maturation regulator, participates in congenital hypogonadotropic hypogonadismPubmed: 32535046
  • Pathophysiological Changes in Female Rats with Estrous Cycle Disorder Induced by Long-Term Heat StressPubmed: 32685488
  • Zihuai recipe alleviates cyclophosphamide-induced diminished ovarian reserve via suppressing PI3K/AKT-mediated apoptosis33422655
  • Zika virus infection in the ovary induces continuously elevated progesterone level and compromises conception in interferon α/β receptor-deficient mice34730391

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