Table 1.

Comparison of experimental approaches for characterization of disulfide-trapped receptor:chemokine complexes

	Amount of sample needed (μg)	Optimal protein concentration (μg/ul)	Preparation time(a)	Throughput(b)	Specificity(c)	Sensitivity(c)
Flow cytometry	n/a (performed on cells)		~1h	+++	−	+++
SDS-PAGE	5–10	0.3–0.5	~1h	++	++	++
Western blotting	0.5–1	0.1–0.5	~4–6 h	++	++	+++
CPM-DSF	0.2–0.4	0.2–0.4	~1h	+++	+++	−
SEC	5–10	0.3–0.5	~0.5–1h	+	+++	−

^(a)The estimated preparation time does not include protein purification. Protein purification is required for all experiments except flow cytometry and, in some implementations, Western blotting, and typically takes at least 3 days.

^(b)Number of samples that can be reasonably tested in parallel in a single experiment, including controls: up to 5–6 (+), up to 12–15 (++), up to 48–96 (+++)

^(c)A qualitative estimate of specificity and sensitivity of each assay towards strong crosslinks favorably compatible with the native complex geometry.