Multiplex Assay Kit for Beclin 1 (BECN1) ,etc. by FLIA (Flow Luminescence Immunoassay)

ATG6; VPS30; Coiled-Coil,Myosin-Like BCL2 Interacting Protein; ATG6 Autophagy Related 6 Homolog

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Beclin 1 (BECN1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Beclin 1 (BECN1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Beclin 1 (BECN1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Beclin 1 (BECN1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-104 97
EDTA plasma(n=5) 97-105 101
heparin plasma(n=5) 94-103 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Beclin 1 (BECN1) ,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 Beclin 1 (BECN1) ,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 Beclin 1 (BECN1) ,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) 80-96% 98-105% 84-92% 87-101%
EDTA plasma(n=5) 79-101% 91-99% 85-95% 94-101%
heparin plasma(n=5) 97-105% 80-103% 81-88% 81-93%

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#:BECN1) 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
Rejuvenation Res. Can Enhanced Autophagy be Associated with Human Longevity? Serum Levels of the Autophagy Biomarker Beclin-1 are Increased in Healthy Centenarians Pubmed:25307612
J Cosmet Dermatol A peel-off facial mask comprising myoinositol and trehalose-loaded liposomes improves adult female acne by reducing local hyperandrogenism and activating autophagy pubmed:28342238
Tumor Biology The novel CXCR4 antagonist, PRX177561, reduces tumor cell proliferation and accelerates cancer stem cell differentiation in glioblastoma preclinical models pubmed:28639900
19 Beclin1 circulating levels and accelerated ageing markers in COPD  DOI: 10.1183/1393003.congress-2017.PA1018
Cell Death & Disease Beclin1 circulating levels and accelerated aging markers in COPD Pubmed:29402890
Clinica Chimica Acta A novel combined score of biomarkers in sputum may be an indicator for lung cancer: A pilot study Doi: 10.1016/j.cca.2018.09.027
International Journal of Clinical and Experimental Pathology Serum concentration of autophagy-related protein 7 is correlated with non-alcoholic fatty liver disease in Chinese adults
EXPERIMENTAL AND THERAPEUTIC MEDICINE Crosstalk between vitamin D axis, inflammation and host immunity mechanisms: A prospective study 33936265
International Journal for Research in Applied Sciences and Biotechnology Inhibition of HepG-2 Cells (Liver Cancer Cell Line) Viability by 3-Hydroxypyridine-2-Carboxaldehyde N (4)-Methylthiosemicarbazone
BIOMARKERS OF AUTOPHAGY AND ANTIVIRAL ROLE OF PLACENTA FOR PREDICTING PRETERM DELIVERY IN PATIENTS WITH PRETERM DELIVERY?¡­
European Journal of Natural History IDENTIFICATION OF FACTORS CONNECTED WITH AUTOPHAGY ACTIVITY UNDER CONDITIONS OF PARTIAL NUTRITIONAL DEPRIVATION IN MEN
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