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Multiplex Assay Kit for High Molecular Weight Kininogen (HMWK) ,etc. by FLIA (Flow Luminescence Immunoassay)
Williams-Fitzgerald-Flaujeac Factor; Fitzgerald Factor; HMWK-Kallikrein Factor
(Note: Up to 8-plex in one testing reaction)
- Product No.LMB805Hu
- Organism SpeciesHomo sapiens (Human) Same name, Different species.
- Sample Typeserum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- Test MethodDouble-antibody Sandwich
- Assay Length4h
- Detection Range0.2-200ng/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.067 ng/mL.
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Result
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Specificity
This assay has high sensitivity and excellent specificity for detection of High Molecular Weight Kininogen (HMWK).
No significant cross-reactivity or interference between High Molecular Weight Kininogen (HMWK) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant High Molecular Weight Kininogen (HMWK) and the recovery rates were calculated by comparing the measured value to the expected amount of High Molecular Weight Kininogen (HMWK) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 85-93 | 90 |
| EDTA plasma(n=5) | 96-105 | 99 |
| heparin plasma(n=5) | 84-105 | 97 |
| sodium citrate plasma(n=5) | 95-104 | 101 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Molecular Weight Kininogen (HMWK) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Molecular Weight Kininogen (HMWK) 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 High Molecular Weight Kininogen (HMWK) 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) | 95-102% | 99-105% | 84-92% | 96-105% |
| EDTA plasma(n=5) | 97-104% | 90-102% | 79-101% | 97-105% |
| heparin plasma(n=5) | 93-101% | 78-103% | 90-97% | 80-93% |
| sodium citrate plasma(n=5) | 80-93% | 97-104% | 96-103% | 99-105% |
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#:HMWK) | 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. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.
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| Magazine | Citations |
| Thromb Res | Factor XII–mediated contact activation related to poor prognosis in disseminated intravascular coagulation PubMed: 26706311 |
| Thrombosis Research | Factor XII-mediated contact activation related to poor prognosis in disseminated intravascular coaCavia (Guinea pig )lation Pubmed:26706311 |
| european journal of endocrinology | Contact system activation and high thrombin generation in hyperthyroidism. pubmed:28137735 |
| J Thromb Thrombolysis | Contact system activation in disseminated intravascular coagulation: activities of prekallikrein and high-molecular-weight kininogen are significant risk factors Pubmed:34993714 |