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Carboxy Methyl Lysine (CML) ELISA Kit ELISA Kit

CML Reactivity: Various Species Colorimetric Competition ELISA 61.7 ng/mL - 5000 ng/mL Plasma, Serum
Pubmed (12)
Catalog No. ABIN6954344
Plus shipping costs $45.00
96 tests
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  • Target
    Carboxy Methyl Lysine (CML)
    • 4
    • 3
    • 3
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Various Species
    Detection Method
    Method Type
    Competition ELISA
    Detection Range
    61.7 ng/mL - 5000 ng/mL
    Minimum Detection Limit
    61.7 ng/mL
    The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of CML in serum, plasma.
    Sample Type
    Plasma, Serum
    Analytical Method
    This assay has high sensitivity and excellent specificity for detection of Carboxymethyl Lysine (CML)
    25.9 ng/mL
    • Pre-coated, ready to use 96-well strip plate, flat buttom
    • Plate sealer for 96 wells
    • Reference Standard
    • Standard Diluent
    • Detection Reagent A
    • Detection Reagent B
    • Assay Diluent A
    • Assay Diluent B
    • Reagent Diluent
    • TMB Substrate
    • Stop Solution
    • Wash Buffer (30 x concentrate)
    • Instruction manual
  • Comment

    Information on standard material:
    The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

    Information on reagents:
    The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

    Information on antibodies:
    The provided antibodies and their host vary in different kits.

    Sample Volume
    50 μL
    Assay Time
    2 h
    1. Prepare all reagents, samples and standards,
    2. Add 50μL standard or sample to each well.
      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.
    Reagent Preparation
    1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
    2. Standard - Reconstitute the Standard with 0.7 mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 5,000 ng/mL. Please prepare 5 tubes containing 0.6 mL Standard Diluent and produce a triple dilution series according to the picture shown below. Mix each tube thoroughly before the next transfer. Set up 5 points of diluted standard such as 5,000 ng/mL, 1,666.7 ng/mL, 555.6 ng/mL, 185.2 ng/mL, 61.7 ng/mL, and the last EP tubes with Standard Diluent is the blank as 0 ng/mL.
    3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, kept for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
    4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
    5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.


    1. The standard for this kit is liquid. Due to its small volume, maybe invisible to the eye.
    2. Making serial dilution in the wells directly is not permitted.
    3. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
    4. Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
    5. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
    6. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
    7. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
    8. Contaminated water or container for reagent preparation will influence the detection result.
    Sample Preparation
    • It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
    • If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
    • If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
    • Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).
    Assay Precision
    Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
    CV(%) = SD/meanX100
    Intra-Assay: CV < 10%
    Inter-Assay: CV < 12%
    For Research Use only
  • Precaution of Use
    The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
    4 °C/-20 °C
    Storage Comment
    1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
    2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.
    Expiry Date
    6 months
  • Zhao, Liu, Ten, Zhang, Yuan, Yu, An: "Plasma heparanase is associated with blood glucose levels but not urinary microalbumin excretion in type 2 diabetic nephropathy at the early stage." in: Renal failure, Vol. 39, Issue 1, pp. 698-701, 2018 (PubMed).

    Liu, Wang, Qi, Zou, Sun: "Antiglycation and antioxidant activities of mogroside extract from Siraitia grosvenorii (Swingle) fruits." in: Journal of food science and technology, Vol. 55, Issue 5, pp. 1880-1888, 2018 (PubMed).

    Di Pino, Currenti, Urbano, Scicali, Piro, Purrello, Rabuazzo: "High intake of dietary advanced glycation end-products is associated with increased arterial stiffness and inflammation in subjects with type 2 diabetes." in: Nutrition, metabolism, and cardiovascular diseases : NMCD, Vol. 27, Issue 11, pp. 978-984, 2017 (PubMed).

    Fang, Wang, Li, Guo et al.: "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 ..." in: Geriatrics & gerontology international, Vol. 16, Issue 4, pp. 522-7, 2016 (PubMed).

    Mishra, Saxena, Shukla, Singh, Meyer, Kruzliak, Khanna: "Association of serum N(ε)-Carboxy methyl lysine with severity of diabetic retinopathy." in: Journal of diabetes and its complications, Vol. 30, Issue 3, pp. 511-7, 2016 (PubMed).

    Mishra, Saxena, Ruia, Prasad, Singh, Khanna, Staffa, Gaspar, Kruzliak: "Increased levels of N(ε)- Carboxy methyl lysine (N(ε)-CML) are associated with topographic alterations in retinal pigment epithelium: A preliminary study." in: Journal of diabetes and its complications, Vol. 30, Issue 5, pp. 868-72, 2016 (PubMed).

    Zhang, Fu, Xu, Niu, An: "Hyperoside reduces albuminuria in diabetic nephropathy at the early stage through ameliorating renal damage and podocyte injury." in: Journal of natural medicines, Vol. 70, Issue 4, pp. 740-8, 2016 (PubMed).

    Yeh, Yang, Pai, Wu, Chen: "Long-term administration of advanced glycation end-product stimulates the activation of NLRP3 inflammasome and sparking the development of renal injury." in: The Journal of nutritional biochemistry, Vol. 39, pp. 68-76, 2016 (PubMed).

    Wu, Li, Li, Sui, Yang, Dong, Xie, Sun: "Inhibition of Advanced Glycation Endproduct Formation by Lotus Seedpod Oligomeric Procyanidins through RAGE-MAPK Signaling and NF-κB Activation in High-Fat-Diet Rats." in: Journal of agricultural and food chemistry, Vol. 63, Issue 31, pp. 6989-98, 2015 (PubMed).

    Noori, Tabibi, Hosseinpanah, Hedayati, Nafar: "Effects of combined lipoic acid and pyridoxine on albuminuria, advanced glycation end-products, and blood pressure in diabetic nephropathy." in: International journal for vitamin and nutrition research. Internationale Zeitschrift für Vitamin- und Ernährungsforschung. Journal international de vitaminologie et de nutrition, Vol. 83, Issue 2, pp. 77-85, 2014 (PubMed).

    Shi, Yu, Yang, Wu: "Advanced glycation end products induce human corneal epithelial cells apoptosis through generation of reactive oxygen species and activation of JNK and p38 MAPK pathways." in: PLoS ONE, Vol. 8, Issue 6, pp. e66781, 2013 (PubMed).

    An, Zhao, Yu, Liu, Gu, Gao: "Plasma sRAGE is independently associated with high sensitivity C-reactive protein in type 2 diabetes without coronary artery disease." in: Diabetes research and clinical practice, Vol. 87, Issue 3, pp. e19-22, 2010 (PubMed).

  • Target
    Carboxy Methyl Lysine (CML)
    Alternative Name
    Carboxymethyl Lysine (CML) (CML ELISA Kit Abstract)
    ALL, BCR1, CML, D22S11, D22S662, PHL, BCR, RhoGEF and GTPase activating protein, BCR
    Target Type
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