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User Manual

ELK05J0519, Carboxymethyllysine (CML)

FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
CML ELISA Kit instruction
Intended use
This CML ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or
therapeutic procedures.The Stop Solution changes the color from blue to yellow and the intensity of the
color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of CML
in the sample, this CML ELISA Kit includes a set of calibration standards. The calibration standards are
assayed at the same time as the samples and allow the operator to produce a standard curve of Optical
Density versus CML concentration. The concentration of CML in the samples is then determined by
comparing the O.D. of the samples to the standard curve.
Sample collection and storages

1. Can’t detect the samples which contain NaN3, because NaN3 inhibits HRP activity of the horseradish
   peroxidase.
2. Extract as soon as possible after Specimen collection, Extracted according to the relevant literature.
   Cell culture supernates and plant exact fluids - Remove particulates by centrifugation and assay
   immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw.
   Materials required but not supplied
3. Standard microplate reader(450nm)
4. Precision pipettes and Disposable pipette tips.
5. 37 ℃incubator
   Precautions
6. Do not substitute reagents from one kit to another. Standard, conjugate and microplates are
   matched for optimal performance. Use only the reagents supplied by manufacturer.
7. Do not remove microplate from the storage bag until needed. Unused strips should be stored at
   2-8°C in their pouch with the desiccant provided.
8. Mix all reagents before using.
   Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25°C)
   Materials supplied
   Name 96 determinations 48 determinations
   Microelisa stripplate 8*12strips 8*6strips
   Standard 0.3ml*6tubes 0.3ml*6tubes
   Sample Diluent 6.0ml 3.0ml
   HRP-Conjugate reagent 6.0ml 3.0ml
   20X Wash solution 25ml 15ml
   Chromogen Solution A 6.0ml 3.0ml
   Chromogen Solution B 6.0ml 3.0ml
   Stop Solution 6.0ml 3.0ml
   Closure plate
   membrane 2 2
   User manual 1 1
   Sealed bags 1 1
   Note: Standard (S0 ￫S5) concentration was followed by:0､312.5､625､1250､2500､5000 pg/mL
   Reagent preparation
   20×wash solution:Dilute with Distilled or deionized water 1:20.
   Assay procedure
9. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.
10. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.
11. Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing sample well;
12. Add 50μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C.
13. Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher.
    Storage: 2-8℃. Validity:six months.
    FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE
    READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!