ELISA Kit for Anti-Gonadotropin Releasing Hormone Antibody (Anti-GnRH)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Anti-Gonadotropin Releasing Hormone Antibody (Anti-GnRH).
No significant cross-reactivity or interference between Anti-Gonadotropin Releasing Hormone Antibody (Anti-GnRH) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Anti-Gonadotropin Releasing Hormone Antibody (Anti-GnRH) and the recovery rates were calculated by comparing the measured value to the expected amount of Anti-Gonadotropin Releasing Hormone Antibody (Anti-GnRH) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 101
EDTA plasma(n=5) 80-99 87
heparin plasma(n=5) 87-101 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Anti-Gonadotropin Releasing Hormone Antibody (Anti-GnRH) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Anti-Gonadotropin Releasing Hormone Antibody (Anti-GnRH) 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 Anti-Gonadotropin Releasing Hormone Antibody (Anti-GnRH) 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) 79-92% 88-95% 97-105% 80-95%
EDTA plasma(n=5) 79-102% 90-97% 79-99% 78-98%
heparin plasma(n=5) 82-97% 80-93% 98-105% 88-95%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 5 times;
5. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
6. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Mol Pharmacol. Maternal exposure to dioxin imprints sexual immaturity of the pups through fixing the status of the reduced expression of hypothalamic gonadotropin-releasing … Pubmed:24132183
Journal of Sports Science &amp; Medicine 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 Males Pubmed:24570624
Environmental Research Perfluorooctane sulfonate effects on the reproductive axis in adult male rats Pubmed:25171141
Toxicology Letters Possible role of serotonin and neuropeptide Y on the disruption of the reproductive axis activity by perfluorooctane sulfonate Pubmed:25623392
Toxicol Sci 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 ovulation PubMed: 26358002
Int J Mol Sci. Overexpression of PRL7D1 in Leydig Cells Causes Male Reproductive Dysfunction in Mice Pubmed:26771609
Oral Presentations OP0019-HPR Effectiveness of A Progressive Resistance Strength Program on Hand Osteoarthritis: A Randomised Controlled Trial 60.1.abstract
Oral Presentations OP0021-HPR Sleep Disorders in Patients with Rheumatoid Arthritis and Relationship with Disease Activity 60.3.abstract
Toxicological Sciences Exposure of pregnant mice to perfluorobutanesulfonate causes hypothyroxinemia and developmental abnormalities in female offspring pubmed:27803384
Chronobiology of the immune reponse OP0020 The Association of Gonadotropin-Releasing Hormone and Cytokines in Rheumatoid Arthritis content:75
Environmental Science & Technology Pyrethroid Insecticide Cypermethrin Accelerates Pubertal Onset in Male Mice via Disrupting Hypothalamic-Pituitary-Gonadal Axis pubmed:28731686
Toxicological Sciences Impact of Perfluorooctane Sulfonate on Reproductive Ability of Female Mice through Suppression of Estrogen Receptor a-Activated Kisspeptin Neurons 10.1093:toxsci
GENES Nectin-like molecule 2, a necessary sexual maturation regulator, participates in congenital hypogonadotropic hypogonadism Pubmed: 32535046
BioMed Research International Pathophysiological Changes in Female Rats with Estrous Cycle Disorder Induced by Long-Term Heat Stress Pubmed: 32685488
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