ELISA Kit for Advanced Glycation End Product (AGE)

AGEs; Advanced Glycation End Products

Specificity

This assay has high sensitivity and excellent specificity for detection of Advanced Glycation End Product (AGE).
No significant cross-reactivity or interference between Advanced Glycation End Product (AGE) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Advanced Glycation End Product (AGE) and the recovery rates were calculated by comparing the measured value to the expected amount of Advanced Glycation End Product (AGE) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-93 89
EDTA plasma(n=5) 85-95 91
heparin plasma(n=5) 87-103 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Advanced Glycation End Product (AGE) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Advanced Glycation End Product (AGE) 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 Advanced Glycation End Product (AGE) 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) 80-101% 79-98% 98-105% 90-102%
EDTA plasma(n=5) 99-105% 80-101% 81-90% 95-104%
heparin plasma(n=5) 82-98% 89-101% 87-103% 80-95%

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
Human Immunology Threshold serum concentrations of tumour necrosis factor alpha (TNFα) as a potential marker of the presence of microangiopathy in children and adolescents with type 1 diabetes mellitus (T1DM) PubMed: S0198885912005642
Experimental Dermatology A long-standing hyperglycaemic condition impairs skin barrier by accelerating skin ageing process PubMed: 22017743
J Hum Nutr Diet. Daily intake of vitamin D- or calcium-vitamin D-fortified Persian yogurt drink (doogh) attenuates diabetes-induced oxidative stress: evidence for antioxidative properties of vitamin D. Pubmed: 23829785
Gene Association of RAGE gene polymorphism with circulating AGEs level and paraoxonase activity in relation to macro-vascular complications in Indian type 2 diabetes mellitus patients. Pubmed: 23721855
Gynecol Endocrinol. Lipoprotein-associated phospholipase A2 and AGEs are associated with cardiovascular risk factors in women with history of gestational diabetes mellitus. Pubmed:24397392
Microvasc Res. Role of advanced glycation end product (AGE)-induced receptor (RAGE) expression in diabetic vascular complications Pubmed:24984291
Free Radic Res. Nitroxides prevent protein glycoxidation in vitro. Pubmed:25363554
Sleep and Breathing Serum advanced glycation end products are associated with insulin resistance in male nondiabetic patients with obstructive sleep apnea Pubmed:25566941
Chem Biol Interact Ascorbic acid and protein glycation PubMed: 26163454
Osteoarthritis Cartilage Establishment of a rabbit model to study the influence of advanced glycation end products accumulation on osteoarthritis and the protective effect of pioglitazone PubMed: 26321377
journal of human hypertension Relationship between serum levels of endogenous secretory RAGE and blood pressure in male nondiabetic patients with obstructive sleep apnea PubMed: 25994997
Redox Biol. Glycation of bovine serum albumin by ascorbate PubMed: 26202868
J Oral Pathol Med Impact of morbid obesity and bariatric surgery on antioxidant/oxidant balance of the unstimulated and stimulated human saliva PubMed: 26608886
J Agric Food Chem Inhibition of Advanced Glycation Endproduct Formation by Lotus Seedpod Oligomeric Procyanidins through RAGE-MAPK Signaling and NF-κB Activation in High-Fat-Diet Rats. PubMed: 26207852
Kineziologija ADVANCED GLYCATION END PRODUCTS&#039;RESPONSE TO RESISTANCE TRAINING IN POSTMENOPAUSAL WOMEN WITH TYPE II DIABETES 140251:
Food Funct. Effects of edible bird&#039;s nest on hippocampal and cortical neurodegeneration in ovariectomized rats PubMed: 25920003
Molecular and Cellular Endocrinology Beneficial effects of urine-derived stem cells on fibrosis and apoptosis of myocardial, glomerular and bladder cells Pubmed:26952874
Advances in Experimental Medicine and Biology Effects of S-Nitroso-N-Acetyl-Penicillamine (SNAP) on Inflammation, Lung Tissue Apoptosis and iNOS Activity in a Rabbit Model of Acute Lung Injury Pubmed:27334732
Eur J Vasc Endovasc Surg The Receptor for Advanced Glycation End Products (Rage) and Its Ligands in Plasma and InfrainCavia (Guinea pig )inal Bypass Vein Pubmed:26905625
molecular and cellular endocrinology Berberine exerts renoprotective effects by regulating the AGEs-RAGE signaling pathway in mesangial cells during diabetic nephropathy. pubmed:28087385
cellular physiology and biochemistry Inhibition of Methylglyoxal-Induced AGEs/RAGE Expression Contributes to Dermal Protection by N-Acetyl-L-Cysteine. pubmed:28214842
General Physiology and Biophysics Glutathione is the main endogenous inhibitor of protein glycation. pubmed:28218612
Osteoarthritis and Cartilage Establishment of a rabbit model to study the influence of advanced glycation end productsaccumulation on osteoarthritis and the protective effect of pioglitazone. pubmed:26321377
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