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GE Healthcare m tris
( a ) SDS–PAGE analysis of protein purification of the B.c His 6 <t>-TssA</t> Nt1 construct. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell debris. Lane 2, cell-free extract. Lane 3, flowthrough (Ni column). Lane 4, final preparation. ( b ) SDS–PAGE analysis of protein purification of the B.c SeMet MBP-TssA CTD construct. The protein, indicated by the arrow, is shown to be ∼90% pure in the final preparation samples and thus was used for crystallization. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell-free extract. Lane 2, unbound material (amylose column). Lane 3, B.c SeMet MBP-TssA CTD. Lane 4, after MBP cleavage. Lane 5, after gel filtration. Lane 6, final preparation. ( c ) B.c His 6 -TssA Nt1 crystals grown in 0.16 M calcium acetate, 0.08 M sodium cacodylate buffer pH 6.5, 14.4%( w / v ) PEG 8000, 20%( v / v ) glycerol. ( d ) B.c TssA CTD crystals grown in 0.1 M sodium chloride, 0.1 M <t>Tris</t> pH 8.0, 15%( v / v ) ethanol, 5%( v / v ) MPD.
M Tris, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 2 article reviews
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1) Product Images from "TssA from Burkholderia cenocepacia: expression, purification, crystallization and crystallographic analysis"

Article Title: TssA from Burkholderia cenocepacia: expression, purification, crystallization and crystallographic analysis

Journal: Acta Crystallographica. Section F, Structural Biology Communications

doi: 10.1107/S2053230X18009706

( a ) SDS–PAGE analysis of protein purification of the B.c His 6 -TssA Nt1 construct. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell debris. Lane 2, cell-free extract. Lane 3, flowthrough (Ni column). Lane 4, final preparation. ( b ) SDS–PAGE analysis of protein purification of the B.c SeMet MBP-TssA CTD construct. The protein, indicated by the arrow, is shown to be ∼90% pure in the final preparation samples and thus was used for crystallization. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell-free extract. Lane 2, unbound material (amylose column). Lane 3, B.c SeMet MBP-TssA CTD. Lane 4, after MBP cleavage. Lane 5, after gel filtration. Lane 6, final preparation. ( c ) B.c His 6 -TssA Nt1 crystals grown in 0.16 M calcium acetate, 0.08 M sodium cacodylate buffer pH 6.5, 14.4%( w / v ) PEG 8000, 20%( v / v ) glycerol. ( d ) B.c TssA CTD crystals grown in 0.1 M sodium chloride, 0.1 M Tris pH 8.0, 15%( v / v ) ethanol, 5%( v / v ) MPD.
Figure Legend Snippet: ( a ) SDS–PAGE analysis of protein purification of the B.c His 6 -TssA Nt1 construct. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell debris. Lane 2, cell-free extract. Lane 3, flowthrough (Ni column). Lane 4, final preparation. ( b ) SDS–PAGE analysis of protein purification of the B.c SeMet MBP-TssA CTD construct. The protein, indicated by the arrow, is shown to be ∼90% pure in the final preparation samples and thus was used for crystallization. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell-free extract. Lane 2, unbound material (amylose column). Lane 3, B.c SeMet MBP-TssA CTD. Lane 4, after MBP cleavage. Lane 5, after gel filtration. Lane 6, final preparation. ( c ) B.c His 6 -TssA Nt1 crystals grown in 0.16 M calcium acetate, 0.08 M sodium cacodylate buffer pH 6.5, 14.4%( w / v ) PEG 8000, 20%( v / v ) glycerol. ( d ) B.c TssA CTD crystals grown in 0.1 M sodium chloride, 0.1 M Tris pH 8.0, 15%( v / v ) ethanol, 5%( v / v ) MPD.

Techniques Used: SDS Page, Protein Purification, Construct, Marker, Crystallization Assay, Filtration

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Electrophoresis:

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Article Snippet: .. 8 µg of total RNA was separated by electrophoresis and blotted onto Hybond-XL nylon membrane (GE Healthcare, RPN2020S). .. The membrane was incubated in 1x Church buffer (0.25 M NaH2 PO4 ; 1 mM EDTA; 7% SDS; 20 mg/ml salmon sperm DNA; 0.5% BSA) for 2 hrs at 28°C and the denatured radioactive probe was added for incubation overnight.

Agarose Gel Electrophoresis:

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Southern Blot:

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Article Snippet: .. For Southern blot analysis, total genomic DNA was extracted from ssku80 mutant strain 20 and the wild type, digested with either Bam HI or Eco RV and transferred to a Hybond‐XL (Amersham Biosciences, UK) nylon membrane. .. Hybridization was performed at 42 °C under low‐stringency conditions in the presence of ULTRAhyb solution (Ambion, Austin, TX).

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Hybridization:

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    GE Healthcare m tris
    ( a ) SDS–PAGE analysis of protein purification of the B.c His 6 <t>-TssA</t> Nt1 construct. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell debris. Lane 2, cell-free extract. Lane 3, flowthrough (Ni column). Lane 4, final preparation. ( b ) SDS–PAGE analysis of protein purification of the B.c SeMet MBP-TssA CTD construct. The protein, indicated by the arrow, is shown to be ∼90% pure in the final preparation samples and thus was used for crystallization. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell-free extract. Lane 2, unbound material (amylose column). Lane 3, B.c SeMet MBP-TssA CTD. Lane 4, after MBP cleavage. Lane 5, after gel filtration. Lane 6, final preparation. ( c ) B.c His 6 -TssA Nt1 crystals grown in 0.16 M calcium acetate, 0.08 M sodium cacodylate buffer pH 6.5, 14.4%( w / v ) PEG 8000, 20%( v / v ) glycerol. ( d ) B.c TssA CTD crystals grown in 0.1 M sodium chloride, 0.1 M <t>Tris</t> pH 8.0, 15%( v / v ) ethanol, 5%( v / v ) MPD.
    M Tris, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 253 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m tris/product/GE Healthcare
    Average 92 stars, based on 253 article reviews
    Price from $9.99 to $1999.99
    m tris - by Bioz Stars, 2020-09
    92/100 stars
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    ( a ) SDS–PAGE analysis of protein purification of the B.c His 6 -TssA Nt1 construct. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell debris. Lane 2, cell-free extract. Lane 3, flowthrough (Ni column). Lane 4, final preparation. ( b ) SDS–PAGE analysis of protein purification of the B.c SeMet MBP-TssA CTD construct. The protein, indicated by the arrow, is shown to be ∼90% pure in the final preparation samples and thus was used for crystallization. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell-free extract. Lane 2, unbound material (amylose column). Lane 3, B.c SeMet MBP-TssA CTD. Lane 4, after MBP cleavage. Lane 5, after gel filtration. Lane 6, final preparation. ( c ) B.c His 6 -TssA Nt1 crystals grown in 0.16 M calcium acetate, 0.08 M sodium cacodylate buffer pH 6.5, 14.4%( w / v ) PEG 8000, 20%( v / v ) glycerol. ( d ) B.c TssA CTD crystals grown in 0.1 M sodium chloride, 0.1 M Tris pH 8.0, 15%( v / v ) ethanol, 5%( v / v ) MPD.

    Journal: Acta Crystallographica. Section F, Structural Biology Communications

    Article Title: TssA from Burkholderia cenocepacia: expression, purification, crystallization and crystallographic analysis

    doi: 10.1107/S2053230X18009706

    Figure Lengend Snippet: ( a ) SDS–PAGE analysis of protein purification of the B.c His 6 -TssA Nt1 construct. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell debris. Lane 2, cell-free extract. Lane 3, flowthrough (Ni column). Lane 4, final preparation. ( b ) SDS–PAGE analysis of protein purification of the B.c SeMet MBP-TssA CTD construct. The protein, indicated by the arrow, is shown to be ∼90% pure in the final preparation samples and thus was used for crystallization. Lane M , Mark12 marker (labelled in kDa). Lane 1, cell-free extract. Lane 2, unbound material (amylose column). Lane 3, B.c SeMet MBP-TssA CTD. Lane 4, after MBP cleavage. Lane 5, after gel filtration. Lane 6, final preparation. ( c ) B.c His 6 -TssA Nt1 crystals grown in 0.16 M calcium acetate, 0.08 M sodium cacodylate buffer pH 6.5, 14.4%( w / v ) PEG 8000, 20%( v / v ) glycerol. ( d ) B.c TssA CTD crystals grown in 0.1 M sodium chloride, 0.1 M Tris pH 8.0, 15%( v / v ) ethanol, 5%( v / v ) MPD.

    Article Snippet: A B.c His6 -TssA Nt1 construct was overexpressed in E. coli BL21 (DE3) cells (Studier & Moffatt, 1986 ) grown in brain heart infusion broth (Becton Dickinson) at 37°C to an OD600 of 0.5–0.7, whereupon expression was induced by the addition of 1 m M IPTG followed by incubation for a further 2–3 h. B.c His6 -TssA Nt1 was purified from clarified cell lysate in 50 m M Tris pH 8.0, 500 m M NaCl, applied onto a HisTrap HP column (GE Healthcare Life Sciences) and eluted with a linear gradient of imidazole (0–350 m M ) in 50 m M Tris pH 8.0, 500 m M NaCl.

    Techniques: SDS Page, Protein Purification, Construct, Marker, Crystallization Assay, Filtration

    Crystals of the 1C1 Fab–rEphA2 complex. The crystals shown in ( a ) and ( b 3. Crystals grew to dimensions of up to 0.3 × 0.3 × 0.02 mm in hanging drops, in which 1.5 µl 10 mg ml −1 protein complex solution in 50 m M Tris–HCl pH 7.5, 100 m M NaCl was mixed with 1 µl reservoir solution (16% PEG 4000, 0.08 M sodium acetate pH 5.0, 0.16 M ammonium acetate). Under these and similar conditions, only crystals of two different morphologies could be obtained (often in the same drop), namely the so-called ‘stacked plates’ ( a ) and ‘book pages’ ( b ).

    Journal: Acta Crystallographica Section F: Structural Biology and Crystallization Communications

    Article Title: Crystallization and preliminary X-ray diffraction analysis of the complex of a human anti-ephrin type-A receptor 2 antibody fragment and its cognate antigen

    doi: 10.1107/S1744309110015861

    Figure Lengend Snippet: Crystals of the 1C1 Fab–rEphA2 complex. The crystals shown in ( a ) and ( b 3. Crystals grew to dimensions of up to 0.3 × 0.3 × 0.02 mm in hanging drops, in which 1.5 µl 10 mg ml −1 protein complex solution in 50 m M Tris–HCl pH 7.5, 100 m M NaCl was mixed with 1 µl reservoir solution (16% PEG 4000, 0.08 M sodium acetate pH 5.0, 0.16 M ammonium acetate). Under these and similar conditions, only crystals of two different morphologies could be obtained (often in the same drop), namely the so-called ‘stacked plates’ ( a ) and ‘book pages’ ( b ).

    Article Snippet: The eluted protein was directly applied onto a HiTrap Q HP column equilibrated with 50 m M Tris–HCl pH 7.5 and eluted in an NaCl gradient according to the manufacturer’s instructions (GE Healthcare).

    Techniques:

    A mass spectrum showing the covalent modification of apo cutinase by E600. The modification was carried out at pH 8.0 in Tris–HCl buffer; cutinase was pre-incubated with a twofold excess of E600 at room temperature for 60 min. Two peaks

    Journal: Acta Crystallographica Section F: Structural Biology and Crystallization Communications

    Article Title: Crystallization and preliminary X-ray analysis of recombinant Glomerella cingulata cutinase

    doi: 10.1107/S1744309108012086

    Figure Lengend Snippet: A mass spectrum showing the covalent modification of apo cutinase by E600. The modification was carried out at pH 8.0 in Tris–HCl buffer; cutinase was pre-incubated with a twofold excess of E600 at room temperature for 60 min. Two peaks

    Article Snippet: The fractions containing cutinase were desalted against 20 m M Tris–HCl pH 7.5 buffer using a HiTrap desalting column (GE Healthcare Biosciences, Sweden) and subsequently cleaved with recombinant enterokinase (Novagen, Germany) according to the manufacturer’s specifications, leaving an additional linker sequence of seven residues (AMAISDP) at the N-­terminus.

    Techniques: Modification, Incubation