ELISA Kit for Kisspeptin 1 (KISS1)

Metastin; KiSS-1 Metastasis-Suppressor

Specificity

This assay has high sensitivity and excellent specificity for detection of Kisspeptin 1 (KISS1).
No significant cross-reactivity or interference between Kisspeptin 1 (KISS1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Kisspeptin 1 (KISS1) and the recovery rates were calculated by comparing the measured value to the expected amount of Kisspeptin 1 (KISS1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 88-96 91
EDTA plasma(n=5) 96-105 101
heparin plasma(n=5) 93-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Kisspeptin 1 (KISS1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Kisspeptin 1 (KISS1) 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 Kisspeptin 1 (KISS1) 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) 80-90% 92-101% 84-98% 79-89%
EDTA plasma(n=5) 92-101% 82-104% 81-102% 78-104%
heparin plasma(n=5) 79-99% 91-102% 93-101% 91-101%

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
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 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Mol Endocrinol. LIM-homeodomain transcription factor Isl-1 mediates kisspeptin&#039;s effect on insulin secretion in mice. Pubmed:24956377
临床儿科杂志 双酚A 与女童性早熟的关系 Fileup: Pdf
J Pediatr Endocrinol Metab The relation of urinary bisphenol A with kisspeptin in girls diagnosed with central precocious puberty and premature thelarche PubMed: 26565542
Archives of Physiology and Biochemistry Circulating levels of leptin, nesfatin-1 and kisspeptin in postmenopausal obese women Pubmed:27011110
International Journal of Endocrinology The Changes of Serum Leptin and Kisspeptin Levels in Chinese Children and Adolescents in Different Pubertal Stages pubmed:27990162
Journal of Clinical Research in Pediatric Endocrinology Basal Serum Neurokinin B Levels in Differentiating Idiopathic Central Precocious Puberty from Premature Thelarche. pubmed:28008860
BioMed Research International Kisspeptin Protein in Seminal Plasma Is Positively Associated with Semen Quality: Results from the MARHCS Study in Chongqing, China
OBESITY SURGERY Lack of Improvement of Sperm Characteristics in Obese Males After Obesity Surgery Despite the Beneficial Changes Observed in Reproductive Hormones Pubmed: 30806913
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