ELISA Kit for Prekallikrein (PK) Homo sapiens (Human) Sandwich ELISA

KLKB1; KLK3; Fletcher Factor; Kallikrein B,Plasma; Kininogenin; Plasma prekallikrein

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  • ELISA Kit for Prekallikrein (PK) Packages (Simulation)
  • ELISA Kit for Prekallikrein (PK) Packages (Simulation)
  • ELISA Kit for Prekallikrein (PK) Results demonstration
  • SEB801Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
sodium citrate plasma(n=5) 90-98 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Prekallikrein (PK) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Prekallikrein (PK) 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 Prekallikrein (PK) 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
sodium citrate plasma(n=5) 78-90% 97-104% 98-105% 78-101%

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 Prekallikrein (PK)

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 Prekallikrein (PK). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Prekallikrein (PK). 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 Prekallikrein (PK), 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 Prekallikrein (PK) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Altered activities of kininase II, an angiotensin converting enzyme, prekallikrein, and nitric oxide in Kuwaiti patients with type 2 diabetesPubMed: 25964383
  • Factor XII–mediated contact activation related to poor prognosis in disseminated intravascular coagulationPubMed: 26706311
  • Factor XII-mediated contact activation related to poor prognosis in disseminated intravascular coaCavia (Guinea pig )lationPubmed:26706311
  • The First Case of Congenital Prekallikrein Deficiency in Korea With a Novel Pathogenic Variant (c. 1198G> T)Pubmed: 30430790
  • Huaier shows anti©\cancer activities by inhibition of cell growth, migration and energy metabolism in lung cancer through PI3K/AKT/HIF©\1¦Á pathway33377619
  • Contact system activation in disseminated intravascular coagulation: activities of prekallikrein and high-molecular-weight kininogen are significant risk factorsPubmed:34993714

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