ELISA Kit for Alpha-1-Acid Glycoprotein (a1AGP)

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

Specificity

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

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) 97-104 101
EDTA plasma(n=5) 98-105 101
heparin plasma(n=5) 89-98 94

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) 90-102% 97-104% 94-104% 97-105%
EDTA plasma(n=5) 97-105% 94-102% 78-91% 93-102%
heparin plasma(n=5) 78-89% 88-95% 78-94% 98-105%

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 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.

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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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