Magnetic Luminex Assay Kit for E2F Transcription Factor 1 (E2F1)

RBBP3; RBP3; RBAP-1; Retinoblastoma-associated protein 1; Retinoblastoma-binding protein 3; pRB-binding protein E2F-1

  • Magnetic Luminex Assay Kit for E2F Transcription Factor 1 (E2F1) Packages (Simulation)
  • Magnetic Luminex Assay Kit for E2F Transcription Factor 1 (E2F1) Packages (Simulation)
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Specificity of the Magnetic Luminex Assay Kit for E2F Transcription Factor 1 (E2F1)

This assay has high sensitivity and excellent specificity for detection of E2F Transcription Factor 1 (E2F1).
No significant cross-reactivity or interference between E2F Transcription Factor 1 (E2F1) and analogues was observed.

Recovery of the Magnetic Luminex Assay Kit for E2F Transcription Factor 1 (E2F1)

Matrices listed below were spiked with certain level of recombinant E2F Transcription Factor 1 (E2F1) and the recovery rates were calculated by comparing the measured value to the expected amount of E2F Transcription Factor 1 (E2F1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 102
EDTA plasma(n=5) 78-91 83
heparin plasma(n=5) 86-93 89
sodium citrate plasma(n=5) 86-102 93

Precision of the Magnetic Luminex Assay Kit for E2F Transcription Factor 1 (E2F1)

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level E2F Transcription Factor 1 (E2F1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level E2F Transcription Factor 1 (E2F1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity of the Magnetic Luminex Assay Kit for E2F Transcription Factor 1 (E2F1)

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of E2F Transcription Factor 1 (E2F1) 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) 94-103% 81-94% 91-101% 96-105%
EDTA plasma(n=5) 80-97% 80-95% 81-101% 90-98%
heparin plasma(n=5) 86-96% 95-105% 85-104% 96-104%
sodium citrate plasma(n=5) 83-101% 78-93% 82-95% 88-104%

Stability of the Magnetic Luminex Assay Kit for E2F Transcription Factor 1 (E2F1)

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.

Assay procedure summary of the Magnetic Luminex Assay Kit for E2F Transcription Factor 1 (E2F1)

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

Test principle of the Magnetic Luminex Assay Kit for E2F Transcription Factor 1 (E2F1)

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

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