ELISA Kit for Alpha-1-Antitrypsin (a1AT) Canis familiaris; Canine (Dog) Competition ELISA

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

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  • ELISA Kit for Alpha-1-Antitrypsin (a1AT) Packages (Simulation)
  • ELISA Kit for Alpha-1-Antitrypsin (a1AT) Packages (Simulation)
  • ELISA Kit for Alpha-1-Antitrypsin (a1AT) Results demonstration
  • CEB697Ca.jpg Typical Standard Curve
  • Certificate 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) 97-105 102
EDTA plasma(n=5) 90-97 93
heparin plasma(n=5) 90-104 98

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) 88-97% 81-92% 94-104% 92-102%
EDTA plasma(n=5) 91-98% 80-90% 96-103% 82-92%
heparin plasma(n=5) 80-95% 78-99% 88-95% 99-105%

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 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

ELISA Kit for Alpha-1-Antitrypsin (a1AT)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Alpha-1-Antitrypsin (a1AT) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Alpha-1-Antitrypsin (a1AT) and unlabeled Alpha-1-Antitrypsin (a1AT) (Standards or samples) with the pre-coated antibody specific to Alpha-1-Antitrypsin (a1AT). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Alpha-1-Antitrypsin (a1AT) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Alpha-1-Antitrypsin (a1AT) in the sample.

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

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