Fecal sample should be suspended in 1 x Assay buffer (1 g/ml) and vortex for 1 min. Centrifuge at 6000 x g for 5 min and collect supernatant for use in ELISA. If the concentration is out of range of standard curve, Assay Buffer volume should be adjusted accordingly.

Treatment of bronchoalveolar lavage fluid for ELISA assay

1. Add equal volume of 1 x BAL Solution to the bronchoalveolar lavage (BAL).
2. Sonicate for 20 sec or mix thoroughly by a pipette to release the cytokine and centrifuge for 3 min at 3000 x g to collect supernatant for ELISA assay.
3. If sampled from a patient with lung disease or patient who suffered severe inflammatory condition or medication, a serial dilution of the BAL supernatant with 1 x BAL Solution may be needed for appropriate measurement using Standard Curve generated by the ELISA kit.

Treatment of Cerebrospinal Fluid for ELISA assay

1. Add equal volume of 1 x CSF Solution to the Cerebrospinal fluid (CSF) and vortex 30 sec.
2. Centrifuge for 3 min at 3000 x g to collect supernatant for ELISA assay.
3. If sampled from a rheumatoid arthritis patient or patient who suffered severe inflammatory condition or medication, a serial dilution of the CSF supernatant with 1 x CSF Solution may be needed for appropriate measurement using Standard Curve generated by the ELISA kit.

Treatment of Synovial Fluid for ELISA assay

1. Add equal volume of 1 x SF Solution to the synovial fluid (SF).
2. Centrifuge for 3 min at 3000 x g to collect supernatant for ELISA assay.
3. If sampled from a rheumatoid arthritis patient or patient who suffered severe inflammatory condition or medication, a serial dilution of the SF supernatant with 1 x SF Solution may be needed for appropriate measurement using Standard Curve generated by the ELISA kit.

Treatment of Urine Samples for ELISA assay

1. Centrifuge urine sample for 3 min at 3000 x g to collect supernatant for ELISA assay.
2. If sample contains high density of analyte, or sampled from a rheumatoid arthritis patient or patient who suffered severe inflammatory condition or medication, a serial dilution of the supernatant with 1 x Assay Buffer may be needed for appropriate measurement using Standard Curve generated by the ELISA kit.

Treatments of whole blood for ELISA

1. Collect 0.3 ml of whole blood to 0.9 ml of 1 x Assay Buffer in a 1.5 ml microcentrifuge, vortex 1 min and leave it at room temperature for 5 min.
2. Centrifuge at 5000 x g for 3 min to collect the supernatant as sample in the Assay Procedures.