Multiplex Assay Kit for Jagged 1 (JAG1) ,etc. by CBA (Cytometric Bead Array)

CD339; AGS; AHD; AWS; HJ1; JAGL1; Alagille Syndrome; Jagged 1 Protein

Specificity

This assay has high sensitivity and excellent specificity for detection of Jagged 1 (JAG1) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Jagged 1 (JAG1) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Jagged 1 (JAG1) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Jagged 1 (JAG1) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 83-102 87
EDTA plasma(n=5) 82-101 86
heparin plasma(n=5) 95-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Jagged 1 (JAG1) ,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 Jagged 1 (JAG1) ,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 Jagged 1 (JAG1) ,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) 99-105% 89-98% 83-91% 93-103%
EDTA plasma(n=5) 81-93% 82-89% 78-99% 79-94%
heparin plasma(n=5) 79-101% 89-96% 86-103% 88-95%

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
13 Cross-talk between the Toll-like receptor 4 and Notch1 pathways augments the inflammatory response in the interstitial cells of stenotic human aortic valves PubMed: PMC3506187
Eur J Obstet Gynecol Reprod Biol. Profiling of selected angiogenesis-related genes in proliferative eutopic endometrium of women with endometriosis Pubmed: 24188612
PLoS One. Augmented Osteogenic Responses in Human Aortic Valve Cells Exposed to oxLDL and TLR4 Agonist: A Mechanistic Role of Notch1 and NF-κB Interaction Pubmed:Pmc4014478
Biochim Biophys Acta.? Ligation of ICAM-1 on human aortic valve interstitial cells induces the osteogenic response: A critical role of the Notch1-NF-κB pathway in BMP-2 expression Pubmed:25101972
international journal of oral-medical sciences Jagged1 Stimulates Bone Resorption during Orthodontic Tooth Movement Dn: Jlc
Oncology letters MicroRNA‑26b acts as an antioncogene and prognostic factor in cervical cancer
Journal of Biological Chemistry Growth hormone induces Notch1 signaling in podocytes and contributes to proteinuria in diabetic nephropathy Pubmed: 31511328
Cell Death & Disease Growth hormone induces mitotic catastrophe of glomerular podocytes and contributes to proteinuria 33795655
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