BovineBrucella ELISA Kit instruction

Intended use

This Brucella 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 Brucella in the sample, this Brucella 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 cutoff value. The existenceor notof Brucella in the samples is then determined by comparing the O.D. of the samples to the CUT OFF.

Samplecollection and storages

Serum-Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximay 3000×g. Remove serum and assay immediay 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 30minutes 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 immediay or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.

Note: The samples shoule be centrifugated dequay 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

1. 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.

2. 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.

3. Mix all reagents before using.

Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25°C)

Determine the result

1. Test validity: the average of Positive control well≥1.00; the average of Negative control well ≤0.15.

2. Calculate Critical (CUT OFF): Critical= the average of Negative control well + 0.15.

Negative Result: sample OD< Calculate Critical (CUT OFF) is Negative.

Positive Result: sample OD≥ Calculate Critical (CUT OFF) is Positive.

Storage and validity

Storage： 2-8℃.

validity： six months.

FOR RESEARCH USE ONLY;

NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS!

PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!