Multiplex Assay Kit for Proliferating Cell Nuclear Antigen (PCNA) ,etc. by CBA (Cytometric Bead Array)
Cyclin
Overview
Properties
- Product No.CMA591Mi
- Organism SpeciesHomo sapiens (Human), Mus musculus (Mouse), Rattus norvegicus (Rat) Same name, Different species.
- ApplicationsFLIA Kit for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategoryTumor immunityDevelopmental science
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Recovery
Matrices listed below were spiked with certain level of recombinant Proliferating Cell Nuclear Antigen (PCNA) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Proliferating Cell Nuclear Antigen (PCNA) ,etc. by CBA (Cytometric Bead Array) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 94-101 | 97 |
| EDTA plasma(n=5) | 99-105 | 102 |
| heparin plasma(n=5) | 79-94 | 88 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Proliferating Cell Nuclear Antigen (PCNA) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Proliferating Cell Nuclear Antigen (PCNA) ,etc. by CBA (Cytometric Bead Array) 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 Proliferating Cell Nuclear Antigen (PCNA) ,etc. by CBA (Cytometric Bead Array) 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) | 85-92% | 80-99% | 95-104% | 82-95% |
| EDTA plasma(n=5) | 82-89% | 95-102% | 80-104% | 92-101% |
| heparin plasma(n=5) | 88-102% | 93-105% | 89-99% | 78-102% |
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 |
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.
Test principle
Anti-Proliferating Cell Nuclear Antigen (PCNA) ,etc. by CBA (Cytometric Bead Array) antibody was coated on the surface of magnetic beads, to make solid phase carrier. Standard or sample were added to the magnetic beads, respectively, in which Proliferating Cell Nuclear Antigen (PCNA) ,etc. by CBA (Cytometric Bead Array) was bound to the antibody connected to the solid phase carrier, and then biotinylated anti-Proliferating Cell Nuclear Antigen (PCNA) ,etc. by CBA (Cytometric Bead Array) antibodies were added. PE-labeled avidin was added. There is a positive correlation between the concentration of Proliferating Cell Nuclear Antigen (PCNA) ,etc. by CBA (Cytometric Bead Array) in the sample and corresponding value of MFI (Median Fluorescence Intensity). Flow Cytometer was used to determine and calculate the sample concentration.
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Citations
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