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User Manual

ELK09J0172, Bovine Chymase (CMA)

FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
Bovine CMA ELISA Kit instruction
Intended use
This CMA 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 CMA in
the sample, this CMA 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 CMA concentration. The concentration of CMA in the samples is then determined by
comparing the O.D. of the samples to the standard curve.
Sample collection and storages
Serum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for
10 minutes at approximately 3000×g. Remove serum and assay immediately or aliquot and store samples
at -20℃or -80℃.Avoid repeated freeze-thaw cycles.
Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 30 minutes
at 3000×g at 2-8℃within 30 minutes of collection. Store samples at -20℃or -80℃. Avoid repeated
freeze-thaw cycles.
Cell culture supernates and other biological fluids - Remove particulates by centrifugation and
assay immediately or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.
Note: The samples shoule be centrifugated dequately and no hemolysis or granule was
allowed.
Materials required but not supplied

  1. Standard microplate reader(450nm).
  2. Precision pipettes and Disposable pipette tips.
  3. 37 ℃incubator.
    Precautions
  4. 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.
  5. 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.
  6. 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 10.0ml 5.0ml
    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
    Note: Standard (S0 ￫S5) concentration was followed by:0,20,40,80,160,320 pg/mL.
    Reagent preparation
    20×wash solution:Dilute with Distilled or deionized water 1:20.
    Assay procedure
  7. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.
  8. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.
  9. Add Sample: Add testing sample 10μl then add 40μl of Sample Diluent to testing sample wel.
  10. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C.
  11. 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. Complete removal of liquid at each step is essential to good performance. After the last wash, remove
    Storage: 2-8℃. Validity:six months.
    FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE
    BEGINNING!