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Proteins in the Pol Iα peak fraction from the Mono-S column were size-fractionated in a 4 12% NuPAGE Bis Tris gel (MES buffer; Invitrogen), and individual gel slices were digested with trypsin and LysC.
(B) MALDI-TOF MS of the peak fraction of PXL2LRR-peptide shown in (A).
The molecular weight of the peptide from the peak fraction (1207.50) indicated is equivalent to the theoretical weight of CLE42.
The purity of the peak fraction was monitored by Coomassie blue-stained SDS-PAGE.
The peak fraction was estimated to be ∼95% pure, as indicated by Coomassie blue-stained SDS-PAGE.
The peak fraction corresponding to the monodispersed NX1αEC was negatively stained with an equal volume of 2% uranyl acetate for 30 s.
As expected from the antibody profiles, tandem mass spectrometry analysis of tryptic peptides derived from the Muc5ac-enriched peak (fraction 20) and the Muc5b-enriched peak (fraction 29) showed that Muc5ac was present in fraction 20 (2 Muc5ac peptides, no Muc5b peptides) and Muc5b and Muc5ac were present in fraction 29 (8 Muc5b peptides, 3 Muc5ac peptides).
Our data also demonstrate an incremental increase in the lens γ-crystallin peak fraction with the αB-R120G mutation, suggesting that αB-crystallin may affect the expression of γ-crystallin.
Gel filtration chromatography of SW480 cytosol reveals that ABC largely runs as a single peak fraction (Fig. 2A; #42; albumin at fraction #47, not shown), while total β-catenin runs as two peak fractions (Fig. 2A; #36 and 42).
The product peak fraction was then collected and lyophilized.
The peak fraction was used for kinesin labeling.
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