ELISA Kit for Alpha-2-Glycoprotein 1, Zinc Binding (aZGP1)

ZA2G; ZAG

Specificity

This assay has high sensitivity and excellent specificity for detection of Alpha-2-Glycoprotein 1, Zinc Binding (aZGP1).
No significant cross-reactivity or interference between Alpha-2-Glycoprotein 1, Zinc Binding (aZGP1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Alpha-2-Glycoprotein 1, Zinc Binding (aZGP1) and the recovery rates were calculated by comparing the measured value to the expected amount of Alpha-2-Glycoprotein 1, Zinc Binding (aZGP1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-101 95
EDTA plasma(n=5) 91-101 95
heparin plasma(n=5) 85-97 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-2-Glycoprotein 1, Zinc Binding (aZGP1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alpha-2-Glycoprotein 1, Zinc Binding (aZGP1) 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-2-Glycoprotein 1, Zinc Binding (aZGP1) 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) 94-103% 82-97% 78-95% 88-104%
EDTA plasma(n=5) 91-105% 81-96% 86-93% 93-102%
heparin plasma(n=5) 83-105% 86-95% 89-105% 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
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
Journal of Proteomics Identification of plasma Complement C3 as a potential biomarker for neuroblastoma using a quantitative proteomic approach ScienceDirect: S1874391913005472
International journal of endocrinology Serum ZAG Levels Were Associated with eGFR Mild Decrease in T2DM Patients with Diabetic Nephropathy. pubmed:28352283
Quantitative Proteomic Analyses of Tear Fluid in Graves' Orbitopathy ISBN 978-82-8377-114-5
Frontiers in Endocrinology Circulating and Adipose Tissue mRNA Levels of Zinc-α2-Glycoprotein, Leptin, High-Molecular-Weight Adiponectin, and Tumor Necrosis Factor-Alpha in … Pubmed:29755407
International Journal of Endocrinology Serum Zinc-α2-Glycoprotein Levels in Patients with or without Coronary Artery Disease in Chinese North Population Pubmed: 32184822
Frontiers in Endocrinology Low Serum ZAG Levels Correlate With Determinants of the Metabolic Syndrome in Chinese Subjects Pubmed: 32265843
Diabetes Metab Syndr Obes Serum ZAG and Adiponectin Levels Were Closely Related to Obesity and the Metabolically Abnormal Phenotype in Chinese Population Pubmed: 32904655
J Endocrinol Invest A study on serum pro-neurotensin (PNT), furin, and zinc alpha-2-glycoprotein (ZAG) levels in patients with acromegaly Pubmed:35670958
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