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User Manual
ELK05J0214, Conjugated linoleic acid (CLA)
B/B0)
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
CLA ELISA Kit instruction
Intended use
This CLA 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 CLA in the sample, this CLA 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 CLA concentration. The concentration of CLA in
the samples is then determined by comparing the O.D. of the samples to
the standard curve.
Sample collection and storages
- Can’t detect the samples which contain NaN3, because NaN3 inhibits
HRP activity of the horseradish peroxidase.
- 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
- Standard microplate reader(450nm)
- Precision pipettes and Disposable pipette tips.
- 37 ℃incubator
Precautions
- 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.
- 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.
- 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、125、
250、500、1000、2000 pg/mL
Reagent preparation
20×wash solution:Dilute with Distilled or deionized water 1:20.
Assay procedure
- Prepare all reagents before starting assay procedure. It is
recommended that all Standards and Samples be added in duplicate to the
Microelisa Stripplate.
- Add standard: Set Standard wells, testing sample wells. Add standard
50μl to standard well.
- Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to
testing sample well;
- Add 50μl of HRP-conjugate reagent to each well, cover with an
adhesive strip and incubate for 60 minutes at 37°C.
- 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!