ELISA Kit for Kisspeptin 1 (KISS1) Homo sapiens (Human) Sandwich ELISA

Metastin; KiSS-1 Metastasis-Suppressor

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  • ELISA Kit for Kisspeptin 1 (KISS1) Packages (Simulation)
  • ELISA Kit for Kisspeptin 1 (KISS1) Packages (Simulation)
  • ELISA Kit for Kisspeptin 1 (KISS1) Results demonstration
  • SEC559Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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) 91-101 97
EDTA plasma(n=5) 90-97 94
heparin plasma(n=5) 82-97 88

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) 96-104% 80-89% 86-103% 79-94%
EDTA plasma(n=5) 94-101% 92-105% 92-105% 90-98%
heparin plasma(n=5) 86-101% 92-101% 79-93% 82-101%

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 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 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 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Kisspeptin 1 (KISS1)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Kisspeptin 1 (KISS1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Kisspeptin 1 (KISS1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Kisspeptin 1 (KISS1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Kisspeptin 1 (KISS1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • LIM-homeodomain transcription factor Isl-1 mediates kisspeptin&#039;s effect on insulin secretion in mice.Pubmed:24956377
  • 双酚A 与女童性早熟的关系Fileup: Pdf
  • The relation of urinary bisphenol A with kisspeptin in girls diagnosed with central precocious puberty and premature thelarchePubMed: 26565542
  • Circulating levels of leptin, nesfatin-1 and kisspeptin in postmenopausal obese womenPubmed:27011110
  • The Changes of Serum Leptin and Kisspeptin Levels in Chinese Children and Adolescents in Different Pubertal Stagespubmed:27990162
  • Basal Serum Neurokinin B Levels in Differentiating Idiopathic Central Precocious Puberty from Premature Thelarche.pubmed:28008860
  • Kisspeptin Protein in Seminal Plasma Is Positively Associated with Semen Quality: Results from the MARHCS Study in Chongqing, China
  • Lack of Improvement of Sperm Characteristics in Obese Males After Obesity Surgery Despite the Beneficial Changes Observed in Reproductive HormonesPubmed: 30806913
  • Serum Kisspeptin and Its Relation to Metabolic Parameters and Glucose Metabolism in Prepubertal and Pubertal Obese ChildrenPubmed: 33224197
  • KISS1R Signaling Modulates Gonadotropin Sensitivity in Mouse Leydig CellPubmed: 33112765
  • Beyond GnRH, LH and FSH: the role of kisspeptin on hypothalalmic-3 pituitary gonadal (HPG) axis pathology and diagnostic consideration 4
  • Kisspeptin and Hematologic Parameters as Predictive Biomarkers for First-Trimester Abortions34239761
  • Comparison of the Kisspeptin levels in early onset preeclampsia and late-onset preeclampsiaPubmed:35039882
  • Serum kisspeptin, leptin, neuropeptide Y, and neurokinin B levels in adolescents with polycystic ovary syndromePubmed:35170267
  • Differential Expression of Kisspeptin System and Kisspeptin Receptor Trafficking during Spermatozoa Transit in the EpididymisPubmed:35205340

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