ELISA Kit for Glycated Hemoglobin A1c (HbA1c)

Glycosylated Hemoglobin; Hemoglobin A1c; Hb1c; HbAIc; HbAIc

Specificity

This assay has high sensitivity and excellent specificity for detection of Glycated Hemoglobin A1c (HbA1c).
No significant cross-reactivity or interference between Glycated Hemoglobin A1c (HbA1c) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Glycated Hemoglobin A1c (HbA1c) and the recovery rates were calculated by comparing the measured value to the expected amount of Glycated Hemoglobin A1c (HbA1c) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-92 88
EDTA plasma(n=5) 90-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glycated Hemoglobin A1c (HbA1c) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Glycated Hemoglobin A1c (HbA1c) 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 Glycated Hemoglobin A1c (HbA1c) 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) 90-101% 81-102% 87-101% 82-92%
EDTA plasma(n=5) 81-94% 98-105% 86-94% 87-102%

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 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
J Am Coll Nutr The Effect of Vitamin D Supplementation on Glycemic Control and Lipid Profile in Patients with Type 2 Diabetes Mellitus PubMed: 26391639
Journal of the American College of Nutrition The Effect of Vitamin D Supplementation on Glycemic Control and Lipid Profile in Patients with Type 2 Diabetes Mellitus. pubmed:26391639
Journal of Endocrinology and Metabolism Circulating Betatrophin and Hepatocyte Growth Factor in Type 2 Diabetic Patients: Their Relationship With Disease Prognosis 405
Molecular nutrition & food research Chicory inulin ameliorates type 2 diabetes mellitus and suppresses JNK and MAPK pathways in vivo and in vitro pubmed:28105758
33 d-Ribose as a Contributor to Glycated Haemoglobin pubmed:29033370
Journal of Functional Foods Hypoglycemic effect of Hypericum attenuatum Choisy extracts on type 2 diabetes by regulating glucolipid metabolism and modulating gut microbiota
West Indian Medical Journal Plasma Cellular Hypoxia, Mitochondrial Dysfunction, Disease Risk and Prognostic Factors in Type 2 Diabetic Patients in Saudi Arabia.
Journal of Occupational and Environmental Hygiene Stressors, allostatic load, and health outcomes among women hotel housekeepers: A pilot study Pubmed: 30615593
Cell Reports Parabacteroides distasonis Alleviates Obesity and Metabolic Dysfunctions via Production of Succinate and Secondary Bile Acids Pubmed: 30605678
Journal of Agricultural and Food Chemistry Gymnemic acid alleviates type 2 diabetes mellitus and suppresses endoplasmic reticulum stress in vivo and in vitro
Journal of Clinical Medicine 1H Nuclear Magnetic Resonance (NMR)-Based Cerebrospinal Fluid and Plasma Metabolomic Analysis in Type 2 Diabetic Patients and Risk Prediction for Diabetic … Pubmed: 31248127
Food & Function Gymnemic Acid Alleviates Inflammation and Insulin Resistance via PPARδ-and NFκB-mediated pathways in db/db mice.
Obesity Surgery Duodenal-Jejunal Bypass Ameliorates Type 2 Diabetes Mellitus by Activating Insulin Signaling and Improving Glucose Utilization in the Brain Pubmed: 31605365
Acta Diabetologica Modulation of gut microbiota contributes to effects of intensive insulin therapy on intestinal morphological alteration in high-fat-diet-treated mice Pubmed: 31749050
Open Access Macedonian Journal of Medical Sciences Inhibition of Receptor for Advanced Glycation End Products as New Promising Strategy Treatment in Diabetic Retinopathy Pubmed: 32165929
Cell Reports Activation of a Specific Gut Bacteroides-Folate-Liver Axis Benefits for the Alleviation of Nonalcoholic Hepatic Steatosis Pubmed: 32783933
The effects of different intensity endurance and resistance exercise on diabetic-related blood profiles in impaired glucose tolerance mice
Revista Internacional de Andrologia Relation of pharmacopenile duplex ultrasonography parameters and glycated hemoglobin (HbA1c) in diabetic patients with erectile dysfunction 33334711
Advanced Theory and Simulations? Identification of Key Contributive Compounds in a Herbal Medicine: A Novel Mathematic¡ªBiological Evaluation Approach
Dis Markers The Roles of Liver Inflammation and the Insulin Signaling Pathway in PM2. 5 Instillation-Induced Insulin Resistance in Wistar Rats 34745386
Antioxidants Antioxidant Effect of Tyr-Ala Extracted from Zein on INS-1 Cells and Type 2 Diabetes High-Fat-Diet-Induced Mice Pubmed:35740008
P001–Inv. Clínica
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