Multiplex Assay Kit for Alpha-1-Acid Glycoprotein (a1AGP) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Alpha-1-Acid Glycoprotein (a1AGP) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Alpha-1-Acid Glycoprotein (a1AGP) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-94 83
EDTA plasma(n=5) 93-101 97
heparin plasma(n=5) 81-98 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-1-Acid Glycoprotein (a1AGP) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 90-98% 97-104% 91-101% 80-102%
EDTA plasma(n=5) 82-95% 86-95% 80-93% 85-104%
heparin plasma(n=5) 85-92% 87-101% 99-105% 92-99%

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#:a1AGP) 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. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
J Reprod Dev.? An Acute-phase Protein as a Regulator of Sperm Survival in the Bovine Oviduct: Alpha 1-acid-glycoprotein Impairs Neutrophil Phagocytosis of Sperm In Vitro Pubmed:Pmc4219990
Molecular Reproduction and Development 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 vitro Pubmed:25123565
BMC Complementary and Alternative Medicine Identification of plasma protein markers common to patients with malignant tumour and Abnormal Savda in Uighur medicine: a prospective clinical study Pubmed:25652121
PLoS One Gut Microbial Dysbiosis May Predict Diarrhea and Fatigue in Patients Undergoing Pelvic Cancer Radiotherapy: A Pilot Study PubMed: 25955845
Sci Rep.  S100A9 and ORM1 serve as predictors of therapeutic response and prognostic factors in advanced extranodal NK/T cell lymphoma patients treated with pegaspargase/gemcitabine pmc:PMC4810364
Journal of Chromatography A 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 stenosis S0021967317311858
Scientific Reports An autoregressive logistic model to predict the reciprocal effects of oviductal fluid components on in vitro spermophagy by neutrophils in cattle pubmed:28667317
33 Transcriptome Analysis Uncovers a Growth-Promoting Activity of Orosomucoid-1 on Hepatocytes. pubmed:28927749
Brazilian Journal of Otorhinolaryngology Inflammatory markers in palatine tonsils of children with obstructive sleep apnea syndrome Pubmed: 30213594
BioRxiv Proteome-wide characterization and biomarker identification of intracranial aneurysm
The FEBS journal Dramatically changed immune‐related molecules as early diagnostic biomarkers of non‐small cell lung cancer Pubmed: 31482685
PLoS One Effects of vaccination on acute-phase protein response in broiler chicken Pubmed: 32191767
Indian Journal of Animal Sciences 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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