Multiplex Assay Kit for Vimentin (VIM) ,etc. by FLIA (Flow Luminescence Immunoassay)

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Vimentin (VIM) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Vimentin (VIM) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Vimentin (VIM) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Vimentin (VIM) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-103 95
EDTA plasma(n=5) 97-105 101
heparin plasma(n=5) 80-101 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vimentin (VIM) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Vimentin (VIM) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 Vimentin (VIM) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 80-103% 95-104% 82-101% 84-91%
EDTA plasma(n=5) 79-90% 79-103% 80-101% 90-97%
heparin plasma(n=5) 78-92% 99-105% 80-92% 85-101%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:VIM) 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. 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.

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Magazine Citations
Cancer Epidemiology, Biomarkers & Prevention Aberrant Vimentin Methylation Is Characteristic of Breast Cancer pubmed:22315367
Scientific Reports Glyoxalase 1-knockdown in human aortic endothelial cells - effect on the proteome andendothelial function estimates. pubmed:27898103
Journal of Proteomics HtrA3 is a cellular partner of cytoskeleton proteins and TCP1α chaperonin Pubmed:29477555
Journal of Biological Chemistry Growth hormone induces Notch1 signaling in podocytes and contributes to proteinuria in diabetic nephropathy Pubmed: 31511328
American Journal of Transplantation Extracellular vesicles derived from injured vascular tissue promote the formation of tertiary lymphoid structures in vascular allografts Pubmed: 31729155
Lung Is Vimentin the Cause or Effect of Obstructive Sleep Apnea Development? Pubmed: 32088750
Cell Biol Toxicol ZNF471 modulates EMT and functions as methylation regulated tumor suppressor with diagnostic and prognostic significance in cervical cancer 33566221
CELL BIOLOGY AND TOXICOLOGY Metastatic suppression by DOC2B is mediated by inhibition of epithelial-mesenchymal transition and induction of senescence 33758996
Comparison of Vimentin Levels Between Preeclamptic and Normotensive Pregnant Women
Advanced glycation end-products associate with podocytopathy in type II diabetic patients
Non-POU Domain-Containing Octamer-Binding (NONO) Protein Stability Regulated by PIN1 is Crucial for Breast Cancer Tumorigenicity Via the MAPK/β …
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