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Measurement of GM-CSF autoantibody

Pulmonary Alveolar Proteinosis

Diagnostic Criteria for autoimmune PAP

Measurement of GM-CSF autoantibody

GM-CSF auto antibody(anti-GM-CSF antibody, auto antibody against GM-CSF) concentrations are measured in a similar manner to that previously described (ref. 4). Briefly, serum samples are diluted 1:3000 with phosphate buffered saline (PBS) containing 0.1% goat serum and 0.1% Tween 20. Separate 50 ml aliquots of diluted serum and the purified GM-CSF antibody standard are incubated at room temperature for 40 minutes in ELISA plates previously coated overnight at 4°C with 1 μg/ml rhGM-CSF and blocked for 1 hour with 1% bovine serum albumin/PBS. After washing four times with PBS 0.1% Tween 20 (PBST), the antibody captured by rhGM-CSF are detected by peroxidase labeled anti-human IgG Fc γ antibody (Sigma). Tetramethylbenzidine is used as a substrate, the reaction was stopped with 1N H2SO4, and absorbance is measured at 450 nm. All assays are performed in triplicate and the mean results are used.

For further inquiry of GM-CSF antibody testing for research purposes, please contact us at the following email address.

■please contact us : gmcsfab@pap-guide.jp

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