ELISA Kit for Alpha-2-Heremans Schmid Glycoprotein (AHSG)

aHSG; HSGa; A2HS; AHS; FETUA; HSG-A2; Fetuin-A; Alpha 2-HS Glycoprotein; Fetuin A; Alpha-2-Z-Globulin; Ba-alpha-2-Glycoprotein

Specificity

This assay has high sensitivity and excellent specificity for detection of Alpha-2-Heremans Schmid Glycoprotein (AHSG).
No significant cross-reactivity or interference between Alpha-2-Heremans Schmid Glycoprotein (AHSG) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Alpha-2-Heremans Schmid Glycoprotein (AHSG) and the recovery rates were calculated by comparing the measured value to the expected amount of Alpha-2-Heremans Schmid Glycoprotein (AHSG) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-101 98
EDTA plasma(n=5) 79-92 83
heparin plasma(n=5) 92-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-2-Heremans Schmid Glycoprotein (AHSG) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alpha-2-Heremans Schmid Glycoprotein (AHSG) 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-Heremans Schmid Glycoprotein (AHSG) 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) 88-97% 82-97% 89-101% 78-101%
EDTA plasma(n=5) 94-105% 97-104% 79-97% 80-95%
heparin plasma(n=5) 80-105% 79-94% 89-105% 91-99%

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
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 Proteomic analysis reveals oxidative stress response as the main adaptative physiological mechanism in cows under different production systems ScienceDirect: S1874391912002084
Journal of Physiology and Biochemistry Curcumin prevents liver fat accumulation and serum fetuin-A increase in rats fed a high-fat diet PubMed: 23430567
BioMed Research International Serum Fetuin-A Levels Related with Microalbuminuria in Diet-Induced Obese Rats PubMed: PMC3591142
Osteoporosis International Estrogen increases the transcription of human α2-Heremans-Schmid-glycoprotein by an interplay of estrogen receptor α and activator protein-1. Pubmed: 24504099
BMC Pharmacology and Toxicology A novel 11β-hydroxysteroid dehydrogenase type1 inhibitor CNX-010-49 improves hyperglycemia, lipid profile and reduces body weight in diet induced obese C57B6/J mice with a potential to provide cardio protective benefits Pubmed:25098735
Acta Dermatovenerologica Croatica Relationship Between Serum Fetuin-A Levels and Carotid Intima-media Thickness in Turkish Patients with Mild to Moderate Psoriasis. A Case-control Study PubMed: 26476900
Mol Cell Endocrinol Fetuin A promotes lipotoxicity in β cells through the TLR4 signaling pathway and the role of pioglitazone in anti-lipotoxicity. PubMed: 25986658
Ann Dermatol Evaluation of Arterial Stiffness in Patients with Beh PubMed: 26719639
Endocrine Visceral and subcutaneous adipose tissue express and secrete functional alpha2hsglycoprotein (fetuin a) especially in obesity pubmed:27738888
Diabetes Care Targeted Microbiome Intervention by Microencapsulated Delayed-Release Niacin Beneficially Affects Insulin Sensitivity in Humans pubmed:29212824
JOURNAL OF ALZHEIMERS DISEASE Acute Anti-Inflammatory Markers ITIH4 and AHSG in Mice Brain of a Novel Alzheimer's Disease Model Pubmed: 30958362
Journal of Alzheimer's Disease A New Serum Biomarker Set to Detect Mild Cognitive Impairment and Alzheimer's Disease by Peptidome Technology Pubmed: 31771070
Sci Rep Ileal transposition helps to regulate plasma hepatokine levels in obese Zucker (Crl: ZUC (ORL)-Lepr fa) rats 33833309
J Obstet Gynaecol Res Predıctıve value of pregnancy of follıcular fluıd fetuın‐A and‐B levels ın infertıle women after intra‐cytoplasmic sperm injection 34708901
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