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Multiplex Assay Kit for Protein Disulfide Isomerase (PDI) ,etc. by CBA (Cytometric Bead Array)

P4HB; ERBA2L; PDIA1; PO4DB; PROHB, DSI, GIT, PDI, PO4HB, P4Hbeta; Prolyl 4-Hydroxylase Subunit Beta; Procollagen-Proline 2-Oxoglutarate 4-Dioxygenase Beta Polypeptide

Specificity

This assay has high sensitivity and excellent specificity for detection of Protein Disulfide Isomerase (PDI) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Protein Disulfide Isomerase (PDI) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Protein Disulfide Isomerase (PDI) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Protein Disulfide Isomerase (PDI) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-97 89
EDTA plasma(n=5) 89-97 93
heparin plasma(n=5) 82-101 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Protein Disulfide Isomerase (PDI) ,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 Protein Disulfide Isomerase (PDI) ,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 Protein Disulfide Isomerase (PDI) ,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) 92-105% 79-91% 82-101% 94-102%
EDTA plasma(n=5) 97-105% 78-103% 93-102% 94-102%
heparin plasma(n=5) 98-105% 78-98% 79-104% 81-101%

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

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
Chromatographia A Proteomic-Based Approach to Study the Mechanism of Cytotoxicity Induced by Interleukin-1α and Cycloheximide article:10.1007
PLoS One Tunable activation of therapeutic platelet-rich plasma by pulse electric field: Differential effects on clot formation, growth factor release, and platelet … Pubmed: 30256831
CALCIUM CONTROLLED ACTIVATION OF PLATELETS VIA ELECTRICAL STIMULATION
Aging (Albany NY) Endothelial microparticle-associated protein disulfide isomerase increases platelet activation in diabetic coronary heart disease 34285139
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