Multiplex Assay Kit for Transforming Growth Factor Beta 3 (TGFb3) ,etc. by FLIA (Flow Luminescence Immunoassay)

TGF-B3; LAP; Latency-associated peptide

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Transforming Growth Factor Beta 3 (TGFb3) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Transforming Growth Factor Beta 3 (TGFb3) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Transforming Growth Factor Beta 3 (TGFb3) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Transforming Growth Factor Beta 3 (TGFb3) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 101
EDTA plasma(n=5) 88-99 93
heparin plasma(n=5) 98-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Transforming Growth Factor Beta 3 (TGFb3) ,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 Transforming Growth Factor Beta 3 (TGFb3) ,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 Transforming Growth Factor Beta 3 (TGFb3) ,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) 92-101% 95-102% 80-89% 88-97%
EDTA plasma(n=5) 98-105% 88-97% 94-102% 84-103%
heparin plasma(n=5) 82-90% 94-105% 84-98% 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#:TGFb3) 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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PLoS ONE A Combinatorial Relative Mass Value Evaluation of Endogenous Bioactive Proteins in Three-Dimensional Cultured Nucleus Pulposus Cells of Herniated Intervertebral Discs: Identification of Potential Target Proteins for Gene Therapeutic Approaches Plosone: Source
J Reprod Immunol The immunomodulating effect of seminal plasma on T cells PubMed: 25799173
Fertility and Sterility Influence of vitamin D and transforming growth factor β3 serum concentrations, obesity, and family history on the risk for uterine fibroids pubmed:27743697
Eur J Pharm Biopharm New scaffolds encapsulating TGF-β3/BMP-7 combinations driving strong chondrogenic differentiation pubmed:28087378
Transforme edici büyüme faktörü-beta-1 nötralizan antikoru ve transforme edici büyüme faktörü-beta-3'ün trakeal darlık gelişimi üzerine etkileri 10.5606/tgkdc.dergisi.2017.14302
Fertility and Sterility Ulipristal acetate decreases transforming growth factor β3 serum and tumor tissue concentrations in patients with uterine fibroids Pubmed:29525690
Scientific Reports TGF-β concentrations and activity are down-regulated in the aqueous humor of patients with neovascular age-related macular degeneration Pubmed:29795291
Immunology Letters The involvement of multifunctional TGF-β and related cytokines in pathogenesis of endometriosis Pubmed: 30367890
Cell Reports Transforming Growth Factor-β3 Regulates Adipocyte Number in Subcutaneous White Adipose Tissue Pubmed: 30332637
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Temporal TGF-β Supergene Family Signalling Cues Modulating Tissue Morphogenesis: Chondrogenesis within a Muscle Tissue Model? Pubmed: 32660137
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AAPS PharmSciTech Nanoparticles and Nanostructured Films with TGF-β3: Preparation, Characterization, and Efficacy 34378118
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