Multiplex Assay Kit for Alpha-1-Antitrypsin (a1AT) ,etc. by FLIA (Flow Luminescence Immunoassay) Chicken (Gallus) Multiplex 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

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

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  • Multiplex Assay Kit for Alpha-1-Antitrypsin (a1AT) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Alpha-1-Antitrypsin (a1AT) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Alpha-1-Antitrypsin (a1AT) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMB697Ga.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) 92-101 96
EDTA plasma(n=5) 96-105 101
heparin plasma(n=5) 97-105 101
sodium citrate plasma(n=5) 95-103 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) 95-105% 83-101% 95-103% 78-101%
EDTA plasma(n=5) 79-105% 79-90% 80-103% 96-103%
heparin plasma(n=5) 94-101% 92-101% 88-103% 86-93%
sodium citrate plasma(n=5) 86-99% 90-97% 82-98% 83-94%

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#:a1AT) 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. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

Multiplex Assay Kit for Alpha-1-Antitrypsin (a1AT) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Anti-Alpha-1-Antitrypsin (a1AT) antibody was coated on the surface of magnetic beads, to make solid phase carrier. Standard or sample were added to the magnetic beads, respectively, in which Alpha-1-Antitrypsin (a1AT) was bound to the antibody connected to the solid phase carrier, and then biotinylated anti-Alpha-1-Antitrypsin (a1AT) antibodies were added. PE-labeled avidin was added. There is a positive correlation between the concentration of Alpha-1-Antitrypsin (a1AT) in the sample and corresponding value of MFI (Median Fluorescence Intensity).

Giveaways

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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