plastic chromatography column  (Thomson Instrument)

Journal: Nature Communications

Article Title: Plant LHC-like proteins show robust folding and static non-photochemical quenching

doi: 10.1038/s41467-021-27155-1

Figure Lengend Snippet: a A scheme of chimeric Li-ELIP protein; see also Supplementary Fig. for the protein amino acid sequence. b Synechocystis membranes and the purified Li-ELIP were separated on CN-PAGE, 5 and 1.4 μg of Chl were loaded, respectively. The gel was scanned (Scan) and Chl fluorescence (Ch FL) was detected after excitation with blue light. Proteins were further separated by SDS-PAGE in the second dimension and the 2D gel was stained with Coomassie Blue (CBB). c SEC separation of Li-ELIP; Chl absorbance and fluorescence were monitored during chromatography by diode-array and fluorescence detectors (Chl DAD and Chl FLD). Fractions of 0.5 mL were collected, and those shown within the grey rectangle were concentrated and used for pigment analysis. d Extracted pigments were analysed by HPLC; molar stoichiometries of the identified pigments are shown in parentheses. Values represent means of three technical replicates; standard deviations were below 10%. e Absorption spectra of Li-ELIP and ELIP2 proteins as recorded by a DAD during SEC. f CD spectra in the visible region of LIL3 and Li-ELIP proteins. Data were scaled to the CD of the Q y region at 660–690 nm.

Article Snippet: Afterwards, the supernatant with resin was loaded onto an empty plastic chromatography column (Econo-Pac, Bio-Rad) for washing steps.

Techniques: Sequencing, Purification, Clear Native PAGE, Fluorescence, SDS Page, Two-Dimensional Gel Electrophoresis, Staining, Chromatography, Circular Dichroism

Journal: Nature Communications

Article Title: Plant LHC-like proteins show robust folding and static non-photochemical quenching

doi: 10.1038/s41467-021-27155-1

Figure Lengend Snippet: a The purified LIL3 ( ZEP + ) and Li-ELIP ( ZEP + ) protein variants were separated on 2D CN/SDS-PAGE; [1] and [2] indicate monomeric and dimeric forms, respectively. b SEC separation of Li-ELIP; Chl absorbance and fluorescence were monitored during chromatography by diode-array and fluorescence detectors (Chl DAD and Chl FLD). Fractions of 0.5 mL were collected and those shown within the grey rectangle were concentrated and used for pigment analysis. c Extracted pigments were analysed by HPLC; molar stoichiometries of the identified pigments are shown in parentheses. Values represent the means of three technical replicates; standard deviations were below 10%. A probable variant of pigment content per monomeric Li-ELIP protein is shown as an inset. d Absorption spectra of Li-ELIP ( ZEP + ), Li-ELIP, and ELIP2 proteins.

Article Snippet: Afterwards, the supernatant with resin was loaded onto an empty plastic chromatography column (Econo-Pac, Bio-Rad) for washing steps.

Techniques: Purification, SDS Page, Fluorescence, Chromatography, Variant Assay