ELISA Kit for Alpha-1-Acid Glycoprotein (a1AGP) Homo sapiens (Human) Sandwich ELISA

AGP1; a1-AGP; ORM1; OMD 1; Orosomucoid 1

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  • ELISA Kit for Alpha-1-Acid Glycoprotein (a1AGP) Packages (Simulation)
  • ELISA Kit for Alpha-1-Acid Glycoprotein (a1AGP) Packages (Simulation)
  • ELISA Kit for Alpha-1-Acid Glycoprotein (a1AGP) Results demonstration
  • SEA816Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Alpha-1-Acid Glycoprotein (a1AGP) and the recovery rates were calculated by comparing the measured value to the expected amount of Alpha-1-Acid Glycoprotein (a1AGP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-94 83
EDTA plasma(n=5) 78-104 97
heparin plasma(n=5) 86-104 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-1-Acid Glycoprotein (a1AGP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alpha-1-Acid Glycoprotein (a1AGP) 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-Acid Glycoprotein (a1AGP) 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) 80-89% 86-98% 92-99% 82-103%
EDTA plasma(n=5) 85-92% 85-101% 88-97% 95-104%
heparin plasma(n=5) 80-99% 97-104% 88-103% 80-101%

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-Acid Glycoprotein (a1AGP)

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-Acid Glycoprotein (a1AGP). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Alpha-1-Acid Glycoprotein (a1AGP). 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-Acid Glycoprotein (a1AGP), 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-Acid Glycoprotein (a1AGP) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • An Acute-phase Protein as a Regulator of Sperm Survival in the Bovine Oviduct: Alpha 1-acid-glycoprotein Impairs Neutrophil Phagocytosis of Sperm In VitroPubmed:Pmc4219990
  • Evidence for a novel, local acute-phase response in the bovine oviduct: Progesterone and lipopolysaccharide up-regulate alpha 1-acid-glycoprotein expression in epithelial cells in vitroPubmed:25123565
  • Identification of plasma protein markers common to patients with malignant tumour and Abnormal Savda in Uighur medicine: a prospective clinical studyPubmed:25652121
  • Gut Microbial Dysbiosis May Predict Diarrhea and Fatigue in Patients Undergoing Pelvic Cancer Radiotherapy: A Pilot StudyPubMed: 25955845
  • S100A9 and ORM1 serve as predictors of therapeutic response and prognostic factors in advanced extranodal NK/T cell lymphoma patients treated with pegaspargase/gemcitabinepmc:PMC4810364
  • Application of a new procedure for liquid chromatography/mass spectrometry profiling of plasma amino acid-related metabolites and untargeted shotgun proteomics to identify mechanisms and biomarkers of calcific aortic stenosisS0021967317311858
  • An autoregressive logistic model to predict the reciprocal effects of oviductal fluid components on in vitro spermophagy by neutrophils in cattlepubmed:28667317
  • Transcriptome Analysis Uncovers a Growth-Promoting Activity of Orosomucoid-1 on Hepatocytes.pubmed:28927749
  • Inflammatory markers in palatine tonsils of children with obstructive sleep apnea syndromePubmed: 30213594
  • Proteome-wide characterization and biomarker identification of intracranial aneurysm
  • Dramatically changed immune‐related molecules as early diagnostic biomarkers of non‐small cell lung cancerPubmed: 31482685
  • Effects of vaccination on acute-phase protein response in broiler chickenPubmed: 32191767
  • Peripheral concentrations of metabolic and inflammatory indicators during transition period and their relationship with postpartum clinical endometritis in dairy …
  • A New Biomarker Tool for Risk Stratification in ¡°De Novo¡± Acute Heart Failure (OROME)

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