ELISA Kit for Angiotensinogen (AGT)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Angiotensinogen (AGT).
No significant cross-reactivity or interference between Angiotensinogen (AGT) and analogues was observed.

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.

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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
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