ELISA Kit for Angiotensinogen (AGT) Homo sapiens (Human) Sandwich ELISA

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

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  • ELISA Kit for Angiotensinogen (AGT) Packages (Simulation)
  • ELISA Kit for Angiotensinogen (AGT) Packages (Simulation)
  • ELISA Kit for Angiotensinogen (AGT) Results demonstration
  • SEA797Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 88-99 94
EDTA plasma(n=5) 88-95 92
heparin plasma(n=5) 87-103 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiotensinogen (AGT) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Angiotensinogen (AGT) 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) 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) 93-102% 93-101% 98-105% 82-104%
EDTA plasma(n=5) 95-103% 82-93% 92-101% 97-105%
heparin plasma(n=5) 84-98% 88-96% 79-102% 98-105%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Angiotensinogen (AGT)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Angiotensinogen (AGT). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Angiotensinogen (AGT). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Angiotensinogen (AGT), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Angiotensinogen (AGT) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • 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
  • The relation between urinary angiotensinogen and proteinuria in renal AA amyloidosis patientsPubMed: 22320202
  • Urinary angiotensinogen as a marker of intrarenal angiotensin II activity in adolescents with primary hypertensionPubMed: PMC3661928
  • Urinary angiotensinogen as a novel marker of obstructive nephropathy in childrenPubmed: 23772991
  • Association between urinary angiotensinogen, hypertension and proteinuria in pregnant women with preeclampsiaSagepub: Source
  • MP707 CHARACTERISTICS AND OUTCOMES OF CHILDREN WITH AUTOSOMAL RECESSIVE POLYCYSTIC KIDNEY DISEASE COHORT FROM THE ESPN/ERA-EDTA REGISTRYOxfordjournals:Source
  • Relationship between blood pressure varPubMed: 26223347
  • Higher Serum Angiotensinogen Is an Indicator of IgA Vasculitis with Nephritis Revealed by Comparative Proteomes AnalysisPubMed: 26098644
  • Korelasi pengukuran antropometri dengan tekanan darah dan angiotensinogen plasma pada dewasaIndex.Php: Art
  • Relationship between blood pressure variability and renal activity of the renin-angiotensin system.pubmed:26223347
  • Pharmacological evidence: a new therapeutic approach to the treatment of chronic heart failure through SUR2B/Kir6.1 channel in endothelial cells.pubmed:27890915
  • 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 stenosisS0021967317311858
  • Pyrrolidine dithiocarbamate inhibits mouse acute kidney injury induced by diclofenac by targeting oxidative damage, cytokines and NF-κB activityPubmed:30036488
  • Role of (Pro) Renin Receptor in Albumin Overload-Induced Nephropathy in RatsPubmed:29846109
  • Quantitation of Total and Free Thiol Angiotensinogen as a Prognostic Marker for PreeclampsiaPubmed: 31069778
  • Multi-omic signatures of atherogenic dyslipidaemia: pre-clinical target identification and validation in humans33407555
  • Association of Ang-(1¨C7) and des-Arg 9 BK as new biomarkers of obesity and cardiometabolic risk factors in adolescents33568792
  • (Pro) renin receptor antagonist PRO20 attenuates nephrectomy©\induced nephropathy in rats via inhibition of intrarenal RAS and Wnt/¦Â©\catenin signaling34057312
  • A novel strategy to identify candidate diagnostic and prognostic biomarkers for gastric cancer34215253

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