TABLE 1.

K_d values determined for GluA2 constructs by different techniques

Technique	Temperature	GluA2S	GnTI−	EndoH	GluA2L	GluA2S-FAM	EndoH-FAM	EndoH-DyLight
SV	10°C	6.0a [2.1–22]				14.9b [7.6–47]
	20°C	8.3 [2.0–22]	11 [0.8–43]	12c [7.0–30]	5.5 [2.8–43]
				8.3 [5.1–27]
				10 [3.0–20]
FDS-SV	20°C						13d [0.76–203]
							2.5e [0.4–7.8]
							10f [3.6–19]
SE	4°C			160 [83–268]
				284 [159–461]
				30g [ND–260]
	10°C			13h [0.14–50]
				17 [ND–74]
				244h [46–643]
FAI	20°C							10.8 [2.4–29]
								11.3 [ND–68]
								8.43 [ND–50.8]

K_d values are reported in nanomolar units. Errors represent the 95% CI using an automated surface projection method, unless indicated otherwise. 10 independent AUC experiments were performed for GluA2S digested with EndoH, as indicated by replicate entries for the mean and 95% CI. SV, sedimentation velocity with absorbance and interference optics; FDS-SV, sedimentation velocity with fluorescence detection optics; SE, sedimentation equilibrium with absorbance optics; FAI, fluorescence anisotropy.

^a68.3% CI.

^bThe data obtained after FAM labeling of GluA2 led to the highest best-fit value, but the labeling does not significantly affect binding within the 95% CI of this assay.

^cGlobal analysis of the absorbance data at 230 nm shown in Fig. 3 D, with additional data acquired from the same experiment at 210 nm shown in Fig. S2 (top right).

^dAnalysis of FDS-SV data only without hydrodynamic constraints.

^eAnalysis of FDS-SV data only with hydrodynamic constraints.

^fAnalysis of FDS-SV data with one additional single high concentration data point measured by absorbance at 488 nm.

^gAnalysis with low loading concentration and 210-nm detection.

^hAnalysis with oil layer to increase pressure.