OneStep ELISA Kit for Alpha-1-Antitrypsin (a1AT) 
SERPINA1; SPAAT; A1-AT; PI; Serpin Peptidase Inhibitor,Clade A(Alpha-1 Antiproteinase/AntiTrypsin)Member 1; Alpha-1 protease inhibitor; Short peptide from AAT; Serpin A1
Overview
Properties
- Product No.OEB697Mu
- Organism SpeciesMus musculus (Mouse) Same name, Different species.
- ApplicationsEnzyme-linked immunosorbent assay for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategoryEnzyme & KinaseInfection immunityHepatology
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Packages (Simulation)
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Packages (Simulation)
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ISO9001: 2008, ISO13485: 2003 Registered
Recovery
Matrices listed below were spiked with certain level of recombinant Alpha-1-Antitrypsin (a1AT) and the recovery rates were calculated by comparing the measured value to the expected amount of Alpha-1-Antitrypsin (a1AT) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 82-94 | 90 |
| EDTA plasma(n=5) | 81-91 | 85 |
| heparin plasma(n=5) | 94-105 | 97 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-1-Antitrypsin (a1AT) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alpha-1-Antitrypsin (a1AT) 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 Alpha-1-Antitrypsin (a1AT) 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) | 85-104% | 93-105% | 93-105% | 90-105% |
| EDTA plasma(n=5) | 81-96% | 80-101% | 87-96% | 96-103% |
| heparin plasma(n=5) | 86-101% | 92-101% | 79-104% | 85-92% |
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 |
Assay procedure summary
1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well, then add 50µL prepared Detection Reagent A to each well. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
5. Add 50µL Stop Solution. Read at 450nm immediately.
Test principle
The test principle applied in this kit is Sandwich enzyme immunoassay. The microplate provided in this kit has been pre-coated with an antibody specific to Alpha-1-Antitrypsin (a1AT). Standards or samples are then added to the appropriate microplate wells with a HRP-conjugated antibody specific to Alpha-1-Antitrypsin (a1AT). After TMB substrate solution is added, only those wells that contain Alpha-1-Antitrypsin (a1AT) and HRP-conjugated antibody 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 Alpha-1-Antitrypsin (a1AT) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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Citations
- Expression and characterization of recombinant human alpha-antitrypsin in transgenic rice seedPubMed: 23376844
- Hepatic steatosis depresses alpha-1-antitrypsin levels in human and rat acute pancreatitisPubMed: 26634430
- Gut Microbial Dysbiosis May Predict Diarrhea and Fatigue in Patients Undergoing Pelvic Cancer Radiotherapy: A Pilot StudyPubMed: 25955845
- Diagnostic Utility of Biomarkers in COPDPubMed: 26106205
- Early prognostic factors in septic shock cancer patients: a prospective study with a proteomic approachPubmed:29315472
- Urinary Proteomics for the Early Diagnosis of Diabetic Nephropathy in Taiwanese PatientsPubmed: 30486327
- Salivary proteins from dysplastic leukoplakia and oral squamous cell carcinoma and their potential for early detectionPubmed: 31706945
- Systemic Alterations of Immune Response-Related Proteins during Glaucoma Development in the Murine Model DBA/2JPubmed: 32585848
- Prenatal diagnosis of maternal serum from mothers carrying β‐thalassemic fetus34559910