Table I. Mass calibration in BNE using unfixed 3.5–16% acrylamide gels.

Dodecylmaltoside-solubilized complexes from bovine heart mitochondria and soluble proteins of the high molecular weight kit were used as mass markers. Proteins with assigned migration distances were visible as blue bands during BNE. The mass/migration data were used for Fig. 1 and supplemental Fig. S1. Coom., Coomassie binding capability of proteins. Dashes (—) indicate migration that could not be estimated at certain time points.

Marker proteins	Source	pI	Coom.	Mass	Migration after various electrophoresis times
					1.5 h	2.5 h	3.5 h	4.5 h	5.5 h	6.5 h	8.5 h	10.5 h
				kDa	mm
Membrane protein complexes
Monomeric RC I	BHM	<7	Yes	∼1000a	28.5	35.0	39.0	42.0	44	46.0	48.0	49.0
Monomeric complex V	BHM	<7	Yes	597a	31.5	40.5	45.0	49.0	51	53.5	56.0	58.0
Dimeric RC III	BHM	<7	Yes	482a	35.5	45.0	51.0	54.0	57	59.0	61.0	63.0
Monomeric RC IV	BHM	<7	Yes	205a	—	56.0	65.0	71.0	75	79.0	83.0	86.0
Monomeric RC II	BHM	<7	Yes	123a	—	61.0	73.0	80.0	86	91.0	97.0	102.0
High molecular weight kitb
Thyroglobulin dimer	Porcine thyroid	—	Yes	669c	30.0	39.0	43.0	46.0	48	50.0	52.5	54.0
Ferritin	Equine spleen	5.4a; 4.2–4.5d	No	440c; 476a	36.0	48.0	54.0	59.0	62	65.0	69.0	71.0
Catalase	Bovine liver	6.8a; 5.4e	Yes	232c; 239a	—	53.0	63.0	71.0	77	—	—	—
Lactate dehydrogenase	Bovine heart	6.0a; ∼5.0f	No	140c; 146a	—	—	—	—	—	—	—	—
BSA	Bovine serum	5.6a; 4.9d	Yes	66c; 66a	—	—	84.0	97.0	—	—	—	—

^a Calculated for the mature protein using the Swiss-Prot database and the pI/Mw tool from the ExPASy proteomics server (www.expasy.ch).

^b From GE Healthcare.

^c Supplier's data (GE Healthcare).

^d Experimental data from Ref. 26.

^e Experimental data from Ref. 27.

^f Estimated here by IEF using the high molecular weight kit.