Multiplex Assay Kit for Sirtuin 1 (SIRT1) ,etc. by FLIA (Flow Luminescence Immunoassay)

SIR2L1; Silent Mating Type Information Regulation 2 Homolog 1; NAD-Dependent Deacetylase Sirtuin-1; SIR2-like protein 1; SirtT1 75 kDa fragment

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Sirtuin 1 (SIRT1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Sirtuin 1 (SIRT1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Sirtuin 1 (SIRT1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Sirtuin 1 (SIRT1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-101 97
EDTA plasma(n=5) 88-95 91
heparin plasma(n=5) 78-90 82

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Sirtuin 1 (SIRT1) ,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 Sirtuin 1 (SIRT1) ,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 Sirtuin 1 (SIRT1) ,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) 80-102% 79-91% 92-101% 80-94%
EDTA plasma(n=5) 95-104% 93-105% 80-88% 98-105%
heparin plasma(n=5) 78-90% 79-91% 83-96% 86-102%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:SIRT1) 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
PLoS One. SIRT1 Gene Polymorphisms Affect the Protein Expression in Cardiovascular Diseases Plos: Source
PLOS ONE A Novel Role for SIRT-1 in L-Arginine Protection against STZ Induced Myocardial Fibrosis in Rats Pubmed:25501750
Journal of Alzheimer&#039;s Disease Serum miR-206 and miR-132 as Potential Circulating Biomarkers for Mild Cognitive Impairment Pubmed:25589731
Respirology Increases in peripheral SIRT1: A new biological characteristic of asthma PubMed: 26040995
PLOS ONE A Remarkable Age-Related Increase in SIRT1 Protein Expression against Oxidative Stress in Elderly: PubMed: 25785999
Endocrine Changes of serum Mir-217 and the correlation with the severity in type 2 diabetes patients with different stages of diabetic kidney disease. pubmed:27522360
Naunyn Schmiedebergs Arch Pharmacol The potential curative effect of rebamipide in hepatic ischemia/reperfusion injury pubmed:28361247
 International journal of cardiology A sirtuin 1/MMP2 prognostic index for myocardial infarction in patients with advanced coronary artery disease pubmed:28043667
International Journal of Cardiology Serum concentrations and gene expression of sirtuin 1 in healthy and slightly overweight subjects after caloric restriction or resveratrol supplementation: A randomized trial pubmed:28029409
Hepatology International Association of SIRT1 gene polymorphism and its expression for the risk of alcoholic fatty liver disease in the Han population pubmed:29189974
Toxicology and Applied Pharmacology Sildenafil protects against bile duct ligation induced hepaticfibrosis in rats: Potential role for silent information regulator 1 (SIRT1) pubmed:28974454
Chemico-biological Interactions Magnolol protects pancreatic β-cells against methylglyoxal-induced cellular dysfunction pubmed:28919305
Nutrients Gene Expression of Sirtuin-1 and Endogenous Secretory Receptor for Advanced Glycation End Products in Healthy and Slightly Overweight Subjects after … Pubmed:30037068
Experimental Gerontology Specific alterations in the circulating levels of the SIRT1, TLR4, and IL7 proteins in patients with dementia Pubmed:30071285
Kidney and Blood Pressure Research The Kidney Specific Protein myo-Inositol Oxygenase, a Potential Biomarker for Diabetic Nephropathy Pubmed: 30504713
Journal of International Medical Research Serum and follicular fluid levels of sirtuin 1, sirtuin 6, and resveratrol in women undergoing fertilization: an observational, clinical study Pubmed: 30556451
BioMed Research International Centella asiatica Prevents Increase of Hippocampal Tumor Necrosis Factor-Independently of Its Effect on Brain-Derived Neurotrophic Factor in Rat Model of Chronic …
学位论文 Influência do tratamento da doença periodontal na sirtuína 1 e na lectina ligadora de manose em indivíduos com doença arterial coronária
Journal of Physics D: Applied Physics Premature Myocardial Infarction: Genetic Variations in SIRT1 Affect Disease Susceptibility
Journal of Geriatric Oncology Serum biomarkers for predicting overall survival and early mortality in older patients with metastatic solid tumors Pubmed: 30952517
Biomed Research International Prevents Increase of Hippocampal Tumor Necrosis Factor-α Independently of Its Effect on Brain-Derived Neurotrophic Factor in Rat Model of … Pubmed: 30956976
Journal of Diabetes and its Complications Vasohibin-1 in T2DM with different UACR
Clinical Interventions in Aging The Serum Concentration of Anti-Aging Proteins, Sirtuin1 and αKlotho in Patients with End-Stage Kidney Disease on Maintenance Hemodialysis Pubmed: 32214805
Compr Psychiatry Crocin alleviates schizophrenia-like symptoms in rats by upregulating silent information regulator-1 and brain derived neurotrophic factor Pubmed: 33045669
Сибирский федеральный университет Влияние гречневой крупы на сиртуиновую активность у подопытных животных в эксперименте
Journal of Cereal Science Dietary buckwheat enhances sirtuin1 without calorie restriction
Clin Interv Aging Molecular Changes in Circulating microRNAs' Expression and Oxidative Stress in Adults with Mild Cognitive Impairment: A Biochemical and Molecular Study 33447019
Antioxidants The Effect of Repeated Whole-Body Cryotherapy on Sirt1 and Sirt3 Concentrations and Oxidative Status in Older and Young Men Performing Different Levels of?¡­ 33396247
Int Urol Nephrol Evaluation of sirtuin 1 (SIRT1) levels in autosomal dominant polycystic kidney disease 33864594
Neurochem Int Dimethyl fumarate abridged tauo-/amyloidopathy in a D-Galactose/ovariectomy-induced Alzheimer's-like disease: Modulation of AMPK/SIRT-1, AKT/CREB/BDNF, AKT ¡­ 34052296
Toxicol Appl Pharmacol Prasugrel anti-ischemic effect in rats: Modulation of hippocampal SUMO2/3-IкBα/Ubc9 and SIRT-1/miR-22 trajectories 34174262
Food Chem Buckwheat-enriched diet alleviates bisphenol A mediated oxidative stress via modulation of sirtuin 1 and antioxidant status in experimental rats 34763933
Life Sci Novel mechanistic insights towards the repositioning of alogliptin in Parkinson's disease 34774622
Int J Mol Sci Intralesional Infiltrations of Arteriosclerotic Tissue Cells-Free Filtrate Reproduce Vascular Pathology in Healthy Recipient Rats Pubmed:35163435
J Clin Med Effects of Coffee on Sirtuin-1, Homocysteine, and Cholesterol of Healthy Adults: Does the Coffee Powder Matter? Pubmed:35683374
Cell & Bioscience OGT Protects Skeletal Muscle From Damage Through SIRT1 O-GlcNAcylation During Cold Stress
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