bio rad unoq 12 column  (Bio-Rad)

Journal: Journal of Biological Chemistry

Article Title: Dissociation and Aggregation of Calpain in the Presence of Calcium

doi: 10.1074/jbc.m105149200

Figure Lengend Snippet: FIG. 3. Fast Protein Liquid Chromatography separation of cal- pain isoforms on a 1 ml UnoQ (Bio-Rad) quaternary ion ex- change column. The earlier peak contained m-Bam-C115S--DraI-m- 80k/NHis1021k calpain, and the later peak contained C105S-m-80k/21k calpain. The column was run in 50 mM Tris-HCl, pH 7.6, 2 mM EDTA, 0.1% (v/v) Triton X-100, 10 mM 2-mercaptoethanol (buffer A) with a gradient of increasing NaCl concentration. a, a mixture of the two proteins resolved without previous exposure to Ca2. b, a mixture of the two proteins resolved following exposure to 5 mM Ca2 for 30 min, quenching with excess EDTA, and dialysis overnight against buffer A.

Article Snippet: Equal amounts (0.75 mg each) of the two proteins were mixed and resolved by chromatography on a Bio-Rad UnoQ (1 ml) ion exchange column with a gradient of [NaCl] in buffer A.

Techniques: Fast Protein Liquid Chromatography, Concentration Assay