Multiplex Assay Kit for Angiotensinogen (AGT) ,etc. by FLIA (Flow Luminescence Immunoassay)

ANHU; SERPINA8; Angiotensin; Renin Substrate; Serpin Peptidase Inhibitor,Clade A,Member 8(Alpha-1 Antiproteinase,Antitrypsin)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 84-92 88
EDTA plasma(n=5) 83-91 87
heparin plasma(n=5) 84-93 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiotensinogen (AGT) ,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 Angiotensinogen (AGT) ,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 Angiotensinogen (AGT) ,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) 85-93% 78-94% 98-105% 87-102%
EDTA plasma(n=5) 97-105% 85-99% 91-101% 83-94%
heparin plasma(n=5) 92-105% 87-104% 78-92% 82-94%

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#:AGT) 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
Transplant International Elevated urinary angiotensinogen a marker of intrarenal renin angiotensin system in hypertensive renal transplant recipients: does it play a role in development of proteinuria in hypertensive renal transplant patients? Wiley: source
Amyloid The relation between urinary angiotensinogen and proteinuria in renal AA amyloidosis patients PubMed: 22320202
Pediatric Mephrology Urinary angiotensinogen as a marker of intrarenal angiotensin II activity in adolescents with primary hypertension PubMed: PMC3661928
Acta Paediatrica Urinary angiotensinogen as a novel marker of obstructive nephropathy in children Pubmed: 23772991
Journal of the Renin-Angiotensin-Aldosterone System Association between urinary angiotensinogen, hypertension and proteinuria in pregnant women with preeclampsia Sagepub: Source
Pediatric Nephrology MP707 CHARACTERISTICS AND OUTCOMES OF CHILDREN WITH AUTOSOMAL RECESSIVE POLYCYSTIC KIDNEY DISEASE COHORT FROM THE ESPN/ERA-EDTA REGISTRY Oxfordjournals:Source
journal of human hypertension Relationship between blood pressure var PubMed: 26223347
PLoS One Higher Serum Angiotensinogen Is an Indicator of IgA Vasculitis with Nephritis Revealed by Comparative Proteomes Analysis PubMed: 26098644
MKA Korelasi pengukuran antropometri dengan tekanan darah dan angiotensinogen plasma pada dewasa Index.Php: Art
Journal of Human Hypertension Relationship between blood pressure variability and renal activity of the renin-angiotensin system. pubmed:26223347
Acta Pharmacol Sin. Pharmacological evidence: a new therapeutic approach to the treatment of chronic heart failure through SUR2B/Kir6.1 channel in endothelial cells. pubmed:27890915
Journal of Chromatography A Application of a new procedure for liquid chromatography/mass spectrometry profiling of plasma amino acid-related metabolites and untargeted shotgun proteomics to identify mechanisms and biomarkers of calcific aortic stenosis S0021967317311858
Life Sciences Pyrrolidine dithiocarbamate inhibits mouse acute kidney injury induced by diclofenac by targeting oxidative damage, cytokines and NF-κB activity Pubmed:30036488
American Journal of Physiology-Renal Physiology Role of (Pro) Renin Receptor in Albumin Overload-Induced Nephropathy in Rats Pubmed:29846109
Methods in Molecular Biology Quantitation of Total and Free Thiol Angiotensinogen as a Prognostic Marker for Preeclampsia Pubmed: 31069778
J Transl Med Multi-omic signatures of atherogenic dyslipidaemia: pre-clinical target identification and validation in humans 33407555
Hypertens Res Association of Ang-(1¨C7) and des-Arg 9 BK as new biomarkers of obesity and cardiometabolic risk factors in adolescents 33568792
Physiol Rep (Pro) renin receptor antagonist PRO20 attenuates nephrectomy©\induced nephropathy in rats via inhibition of intrarenal RAS and Wnt/¦Â©\catenin signaling 34057312
Cancer Cell Int A novel strategy to identify candidate diagnostic and prognostic biomarkers for gastric cancer 34215253
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