Journal: The Journal of Biological Chemistry

Article Title: Silk Gland Factor-2, Involved in Fibroin Gene Transcription, Consists of LIM Homeodomain, LIM-interacting, and Single-stranded DNA-binding Proteins *

doi: 10.1074/jbc.M113.514471

Figure Lengend Snippet: SGF-2 is a 1.1-MDa heteromeric complex. A , chromatography chart of SGF-2 activity passed through a Bio-Silect 250 gel filtration column. B , EMSA was performed using the E site probe, and each fraction passed through a Bio-Silect 250 column. Source Q fr

Article Snippet: The total eluate (0.15 mg, 2.1 ml) was dialyzed against TEMGTK100 and loaded onto a column of Bio-Silect 250 (Bio-Rad) equilibrated with TEMGTK100 .

Techniques: Multiple Displacement Amplification, Chromatography, Activity Assay, Filtration

Journal: The Journal of Cell Biology

Article Title: Rho- and Rac-dependent Assembly of Focal Adhesion Complexes and Actin Filaments in Permeabilized Fibroblasts: An Essential Role for Ezrin/Radixin/Moesin Proteins

doi:

Figure Lengend Snippet: Purification of the active component from pig brain cytosol. ( A ) Cibachrome blue 3GA: a step-eluted fraction from fast-flow Q-Sepharose, diluted to 50 mM salt, was passed over a 40 ml Cibachrome blue 3GA column, washed extensively, and eluted as shown in a continuous 25-300 μS salt gradient. Protein was monitored in line by absorbence at 280 nm, and salt concentration by conductivity, as shown. The activities of column fractions were assayed and plotted ( far left ordinate axis ) and the two activity peaks were pooled. ( B ) Phenyl–Sepharose: pooled fractions from A were applied to a 15-ml phenyl–Sepharose column and proteins eluted with a 300–25 μS salt gradient. ( C ) Q-Sepharose: pooled active fractions from B were diluted to 20 mM salt and chromatographed on a 5-ml Q-Sepharose column. Activity was eluted in a salt gradient to 200 μS. ( D ) 200 μl of the peak fraction from C was applied to a Bio-Silect gel filtration column with a rated separation range of 100–5 kD. The elution positions of molecular weight standards are indicated.

Article Snippet: For gel filtration, a 200-μl aliquot of the activity peak from the final Q-Sepharose column was applied to a 6-ml Bio-Silect chromatography column (Bio Rad Laboratories: rated fractionation range 100–5 kD) and protein was eluted in DK150, sampling 0.25-ml fractions.

Techniques: Purification, Flow Cytometry, Concentration Assay, Activity Assay, Filtration, Molecular Weight

Journal: Molecular Biology of the Cell

Article Title: Identification of a Novel Leucine-rich Repeat Protein as a Component of Flagellar Radial Spoke in the Ascidian Ciona intestinalis

doi: 10.1091/mbc.02-06-0089

Figure Lengend Snippet: Gel filtration of KI extract by BioSilect SEC400 column. (A) KI extract from KCl-treated axonemes was loaded on a gel filtration column (300 × 7.8 mm) and separated at a flow rate of 1.0 ml/min. Fractions (200 μl) were collected. The numbers above the chromatogram indicate the fraction number. Arrows show the elution positions of molecular mass markers as follows: thyroglobulin (670 kDa), immunoglobulin G (150 kDa), ovalbumin (44 kDa), and myoglobin (17 kDa). (B) Proteins in the fraction 4–25 were separated by 10% SDS-PAGE and immunoblotted by anti-LRR37 and anti-RSP3 antibodies. Both proteins were coeluted at around the position with molecular mass of 1300 kDa.

Article Snippet: The supernatant was separated on a BioSilect SEC400 gel filtration column (300 × 7.8 mm; Bio-Rad , Hercules, CA) at a flow rate of 1.0 ml/min.

Techniques: Filtration, Flow Cytometry, SDS Page