ELISA Kit for Alpha-1-Antichymotrypsin (a1ACT) Homo sapiens (Human) Sandwich ELISA

A1-ACT; SERPINA3; AACT; ACT; GIG24; GIG25; Serpin Peptidase Inhibitor Clade A Member 3(Alpha-1 Antiproteinase; Antitrypsin); Cell growth-inhibiting gene 24/25 protein

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  • ELISA Kit for Alpha-1-Antichymotrypsin (a1ACT) Packages (Simulation)
  • ELISA Kit for Alpha-1-Antichymotrypsin (a1ACT) Packages (Simulation)
  • ELISA Kit for Alpha-1-Antichymotrypsin (a1ACT) Results demonstration
  • SEB015Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Alpha-1-Antichymotrypsin (a1ACT) and the recovery rates were calculated by comparing the measured value to the expected amount of Alpha-1-Antichymotrypsin (a1ACT) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-102 97
EDTA plasma(n=5) 93-101 97
heparin plasma(n=5) 99-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-1-Antichymotrypsin (a1ACT) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alpha-1-Antichymotrypsin (a1ACT) 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-Antichymotrypsin (a1ACT) 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) 92-105% 96-103% 95-104% 78-92%
EDTA plasma(n=5) 82-92% 97-105% 94-101% 84-91%
heparin plasma(n=5) 98-105% 82-93% 84-96% 95-104%

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 Alpha-1-Antichymotrypsin (a1ACT)

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 Alpha-1-Antichymotrypsin (a1ACT). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Alpha-1-Antichymotrypsin (a1ACT). 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 Alpha-1-Antichymotrypsin (a1ACT), 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 Alpha-1-Antichymotrypsin (a1ACT) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Identification of Altered Plasma Proteins by Proteomic Study in Valvular Heart Diseases and the Potential Clinical SignificancePubMed: PMC3754973
  • Identifizierung neuer potentieller Biomarker für das Kolonkarzinom sowie dessen VorstufenInfo:Source
  • 可拋棄式退化性關節炎電化學式尿液免疫感測器之研究 Electrochemical Immunosensor Detection of Urinary Biomaker for Osteoarthritis Diagnosis Publication: U0074-2807201500521300
  • 羊瘙痒因子139A感染引起小鼠脑组织中的α1-ACT表达增加CJFDLAST2017

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