Multiplex Assay Kit for Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) 
N(6)-Carboxymethyllysine
Overview
Properties
- Product No.CMB977Ge
- Organism SpeciesPan-species (General) Same name, Different species.
- ApplicationsFLIA Kit for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategoryMetabolic pathwayEndocrinologyCardiovascular biologyGenetic scienceNutrition metabolism
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Packages (Simulation)
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Packages (Simulation)
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ISO9001: 2008, ISO13485: 2003 Registered
Recovery
Matrices listed below were spiked with certain level of Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 81-97 | 93 |
| EDTA plasma(n=5) | 82-95 | 86 |
| heparin plasma(n=5) | 97-105 | 101 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) 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 Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) 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) | 79-105% | 94-102% | 84-96% | 80-101% |
| EDTA plasma(n=5) | 93-104% | 88-105% | 82-104% | 80-99% |
| heparin plasma(n=5) | 88-97% | 83-94% | 78-101% | 87-101% |
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 |
| 96-well plate | 1 | Plate sealer for 96 wells | 4 |
| Pre-Mixed Standard | 2 | Standard Diluent | 1×20mL |
| Pre-Mixed Magnetic beads (22#:CML) | 1 | Analysis buffer | 1×20mL |
| Pre-Mixed Detection Reagent A | 1×120μL | Assay Diluent A | 1×12mL |
| Detection Reagent B (PE-SA) | 1×120μL | Assay Diluent B | 1×12mL |
| Sheath Fluid | 1×10mL | Wash Buffer (30 × concentrate) | 1×20mL |
| Instruction manual | 1 |
Assay procedure summary
1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 50μL standard or sample to each well, add 50μL prepared Detection Reagent A, add 10μL magnetic beads, and incubate 90 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
5. Wash plate once on magnetic frame;
6. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.
Test principle
Anti-Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) antibody was coated on the surface of magnetic beads. A competitive inhibition reaction is launched between biotin labeled Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) and unlabeled Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) (Standards or samples) with the pre-coated antibody specific to Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array). PE-labeled avidin was added. The amount of bound PE conjugate is reverse proportional to the concentration of Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) in the sample, the intensity of MFI developed is reverse proportional to the concentration of Carboxymethyl Lysine (CML) ,etc. by CBA (Cytometric Bead Array) in the sample.
Giveaways
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Citations
- Advanced Glycation End Products Induce Human Corneal Epithelial Cells Apoptosis through Generation of Reactive Oxygen Species and Activation of JNK and p38 MAPK PathwaysPubMed: PMC3680386
- Effects of combined lipoic acid and pyridoxine on albuminuria, advanced glycation end-products, and blood pressure in diabetic nephropathy.Pubmed: 24491880
- Advanced glycation end-products accelerate the cardiac aging process through the receptor for advanced glycation end-products/transforming growth factor-β-Smad signaling pathway in cardiac fibroblastsPubMed: 26016731
- 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
- Association of serum N Îľ-Carboxy methyl lysine with severity of diabetic retinopathyPubmed:26782022
- Hyperoside reduces albuminuria in diabetic nephropathy at the early stage through ameliorating renal damage and podocyte injury.Pubmed:27255369
- Increased levels of N(ε)- Carboxy methyl lysine (N(ε)-CML) are associated with topographic alterations in retinal pigment epithelium: A preliminary studyPubmed:27039312
- Long-term administration of advanced glycation end-product stimulates the activation of NLRP3 inflammasome and sparking the development of renal injury.pubmed:27816762
- Plasma heparanase is associated with blood glucose levels but not urinary microalbumin excretion in type 2 diabetic nephropathy at the early stage pubmed:28994624
- High intake of dietary advanced glycation end-products is associated with increased arterial stiffness and inflammation in subjects with type 2 diabetes.pubmed:28958695
- Antiglycation and antioxidant activities of mogroside extract from (Swingle) fruitsPubmed:29666541
- Elevated advanced glycation end products are associated with subfoveal ellipsoid zone disruption in diabetic macular edema34708772