Journal: The Journal of Biological Chemistry

Article Title: Structure and reactivity of a siderophore-interacting protein from the marine bacterium Shewanella reveals unanticipated functional versatility

doi: 10.1074/jbc.RA118.005041

Figure Lengend Snippet: Time-resolved formation of the Fe(II)-ferrozine complex by SfSIP. A, experiments performed using ferric-avaroferrin as iron-source and different electron donors. B, experiments performed using NADPH as electron donor with different ferric siderophore.

Article Snippet: The ferrozine assay The ferrozine assay monitored the formation of ferrous iron using an iron chelator with high affinity for Fe 2+ and ferrozine, purchased from Fluka.

Techniques:

Journal: Frontiers in Plant Science

Article Title: Two-dimensional phos-tag zymograms for tracing phosphoproteins by activity in-gel staining

doi: 10.3389/fpls.2015.00230

Figure Lengend Snippet: IEF in-gel activity staining compatibility with Pro-Q Diamond® staining . Separation of phosvitin and protein extracts (50 μg) in non-reducing IEF pH 3–10. (A) Phosvitin separation followed by Pro-Q Diamond® and CCB staining. P, phosphorylated; D, partially dephosphorylated. (B) Separation of protein extracts followed by in-gel activity, Pro-Q Diamond®, and CCB staining. (a) Fe, Fe(III)-citrate/ferrozine/NADH (microsomes, pea roots); (b) TMB, TMB/H 2 O 2 (soluble fraction, corn leaves). Each in-gel activity staining procedure was followed by Pro-Q Diamond® staining in the same gel. Type of staining is indicated at the bottom and pI of the bands indicated on the left of the gels. Cathode and anode are indicated as (−) and (+), respectively. Arrows show the direction of protein separation.

Article Snippet: After reduction, the Fe(II) is bound in a stable complex with ferrozine (Viollier et al., ).

Techniques: Activity Assay, Staining

Journal: Frontiers in Plant Science

Article Title: Two-dimensional phos-tag zymograms for tracing phosphoproteins by activity in-gel staining

doi: 10.3389/fpls.2015.00230

Figure Lengend Snippet: In-gel activity in two-dimensional zymograms . Three different in-gel detections were tested for hrCNE 4–16% and phos-tag hrCNE 4–16%. (A) TMB/H 2 O 2 staining of soluble proteins (50 μg) from maize leaves. (B) NADH-dependent Fe(III)-reductase staining with ferrozine for soluble proteins (75 μg) of pea roots. (C) DAB/H 2 O 2 staining for total protein extracts (50 μg) from corn roots. Separation type for the first dimension (IEF or NEPHGE) is indicated on top and second dimension on the side of the gels. The pH of the IEF (3–10) is indicated on the top of the first dimension. Arrows show the direction of protein separation. Cathode and anode are indicated as (−) and (+), respectively. Numbered spots were analyzed for the shift on phos-tag hrCNE compared to hrCNE and analyzed by LC-MS/MS, numbers of potentially phosphorylated proteins are written in italic letters.

Article Snippet: After reduction, the Fe(II) is bound in a stable complex with ferrozine (Viollier et al., ).

Techniques: Activity Assay, Staining, Liquid Chromatography with Mass Spectroscopy