Table 1 |.

SAA conditions reported in selected publications for the detection of αSyn aggregates in human CSF

Ref.	Biological sample	Substrate (source)	Reaction mix	Reaction volume (μl)	Replicates (CSF volume, μl)	Assay instrument	Assay length (h)	SAA cycle (min)	Shaking	Incubation time (min), temperature (°C)	Bead type (no.)
Fairfoul et al.13	CSFand BH	0.1 mg/ml rec-αSyn WT, no tag (Stratech)	100 mM phosphate buffer pH 8.2, 10 μm ThT	100	2 (15)	OPTIMA FluoSTAR Omega fluorometer	120	15	Double orbital 1 min, 200 rpm	14, 30	0.5-mm zirconium silica (37 mg)
Shahnawaz et al.14 (original αSyn-SAA)	CSF and BH	1 mg/ml rec-αSyn WT+6hist (in house)	100 mM PIPES pH 6.5, 500 mM NaCI, 5 μm ThT	200	3 (40)	Thermomixer R Gemini Spectra EM fluorometer	350–400	30	Orbital 1 min, 500 rpm	29, 37	No beads
Groveman et al.15	CSF and BH	0.1 mg/ml rec-αSyn K23Q +6hist (in house)	40 mM phosphate buffer pH 8, 170 mM NaCI, 10 μm ThT, 0.0015% (m/v) SDS	100	4(15)	FLUOstar Omega fluorometer	50	2	Double orbital 1 min, 400 rpm	1, 42	0.8-mm silica (6)
Bargar et al.57	CSF, brain, skin, salivary gland and colon	1 mg/ml rec-αSyn WT, no tag (rPeptide)	40 mM phosphate buffer pH 8, 170 mM NaCI, 20 μm ThT	100	4(2)	FLUOstar Omega fluorometer	60	2	Double orbital 1 min, 400 rpm	1, 42	0.8-mm silica (6)
Concha-Marambio et al.16 (optimized αSyn-SAA)	CSF	0.3 mg/ml rec-αSyn WT +6hist (in house)	100 mM PIPES pH 6.5, 500 mM NaCI, 5 μm ThT	200	3 (40)	FLUOstar Omega fluorometer	150	30	Double orbital 1 min, 700 rpm	29, 37	3/32-inch Si3N4 bead (1)
Russo et al.17 (Abbvie protocol)	CSF	0.1 mg/ml rec-αSyn WT, no tag (in house)	100 mM phosphate buffer pH 8.2,10 μM ThT	100	8 (5)	Not specified	70	15	Double orbital 1 min, 200 rpm	14, 37	0.10-mm zirconia/silica beads (25 mg)
Perra et al.60	CSF and olfactory mucosa	0.05 mg/ml rec-αSyn WT, no tag (in house)	100 mM phosphate buffer pH 8.2, 10 μm ThT, 0.0075% (m/v) SDS	100	4(15)	FLUOstar Omega fluorometer	80	15	Double orbital 1 min, 200 rpm	14, 30	0.5-mm glass beads (37 mg)