Multiplex Assay Kit for Tubulin Beta (TUBb) ,etc. by CBA (Cytometric Bead Array)

TUB-B; TUBB5; TUBB1; M40

Specificity

This assay has high sensitivity for detection of Tubulin Beta (TUBb) ,etc. by CBA (Cytometric Bead Array).
100% cross-reactivity of Tubulin Beta (TUBb) ,etc. by CBA (Cytometric Bead Array) was observed among .

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 85-99 92
EDTA plasma(n=5) 79-90 84
heparin plasma(n=5) 80-97 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tubulin Beta (TUBb) ,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 Tubulin Beta (TUBb) ,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 Tubulin Beta (TUBb) ,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) 89-96% 90-104% 95-102% 96-104%
EDTA plasma(n=5) 93-101% 84-94% 99-105% 98-105%
heparin plasma(n=5) 95-102% 86-99% 89-99% 93-102%

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.

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Magazine Citations
archives of pharmacal research Synthesis and cytotoxic activity of certain trisubstituted azetidin-2-one derivatives as a cis-restricted combretastatin A-4 analogues pubmed:27747473
archiv der pharmazie (Weinheim). New 3-Substituted-2-(4-hydroxyanilino)pyridine Derivatives: Synthesis, Antitumor Activity, and Tubulin Polymerization Inhibition. pubmed:28150327
ACS Applied Materials & Interfaces Novel Beta-Tubulin-Immobilized Nanoparticles Affinity Material for Screening β-Tubulin Inhibitors from a Complex Mixture pubmed:28112513
Future Medicinal Chemistry Design, synthesis and biological evaluation of chromenopyrimidines as potential cytotoxic agents Pubmed:29779400
Future Medicinal Chemistry Design and synthesis of novel 5-(4-chlorophenyl) furan derivatives with inhibitory activity on tubulin polymerization Pubmed:29966433
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Journal of Enzyme Inhibition and Medicinal Chemistry Design, synthesis, and screening of amino thiophene carboxamide derivatives on hepatocellular carcinomaas VEGFR-2Inhibitors Pubmed: 30191744
European Journal of Medicinal Chemistry Potent combretastatin A-4 analogs containing 1, 2, 4-triazole: Synthesis, antiproliferative, anti-tubulin activity, and docking study Pubmed: 31536891
ACS Sensors Early diagnosis of Alzheimer's disease in blood using a disposable electrochemical microfluidic platform Pubmed: 32207606
Bioorganic Chemistry Tubulin Inhibitors: Discovery of a New Scaffold Targeting Extra-binding Residues within the Colchicine Site through Anchoring Substituents Properly Adapted to their … Pubmed: 32325332
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