Structured Review

GE Healthcare m dtt
Disulfide reduction during catalysis by Ero1p and the de-regulated mutant C150A/C295A. Enzyme at a final concentration of 2 μ M was mixed with 100 μ M <t>Trx</t> red . Aliquots were removed from the reactions at the indicated time points, and disulfide status was preserved by blocking with maleimide reagents. The migration of Ero1p on SDS-PAGE at the various time points was examined and compared with wild-type Ero1p (WT), the indicated Ero1p mutants, and <t>DTT-treated</t> Ero1p (WT reduced). (A) The Trx red oxidation reaction was blocked with AMS and separated by SDS-PAGE under nonreducing conditions. The asterisk (*) indicates a species that migrates slower than Ero1p C90A/C349A when blocked with AMS, but faster than this disulfide mutant when blocked with NEM (compare with panel C). (B) The same experiment as in (A) was performed, but the samples were reduced with DTT after AMS treatment before separation by SDS-PAGE. (C) Reactions were blocked with NEM and applied to the gel under nonreducing conditions.
M Dtt, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Steps in reductive activation of the disulfide-generating enzyme Ero1p"

Article Title: Steps in reductive activation of the disulfide-generating enzyme Ero1p

Journal: Protein Science : A Publication of the Protein Society

doi: 10.1002/pro.473

Disulfide reduction during catalysis by Ero1p and the de-regulated mutant C150A/C295A. Enzyme at a final concentration of 2 μ M was mixed with 100 μ M Trx red . Aliquots were removed from the reactions at the indicated time points, and disulfide status was preserved by blocking with maleimide reagents. The migration of Ero1p on SDS-PAGE at the various time points was examined and compared with wild-type Ero1p (WT), the indicated Ero1p mutants, and DTT-treated Ero1p (WT reduced). (A) The Trx red oxidation reaction was blocked with AMS and separated by SDS-PAGE under nonreducing conditions. The asterisk (*) indicates a species that migrates slower than Ero1p C90A/C349A when blocked with AMS, but faster than this disulfide mutant when blocked with NEM (compare with panel C). (B) The same experiment as in (A) was performed, but the samples were reduced with DTT after AMS treatment before separation by SDS-PAGE. (C) Reactions were blocked with NEM and applied to the gel under nonreducing conditions.
Figure Legend Snippet: Disulfide reduction during catalysis by Ero1p and the de-regulated mutant C150A/C295A. Enzyme at a final concentration of 2 μ M was mixed with 100 μ M Trx red . Aliquots were removed from the reactions at the indicated time points, and disulfide status was preserved by blocking with maleimide reagents. The migration of Ero1p on SDS-PAGE at the various time points was examined and compared with wild-type Ero1p (WT), the indicated Ero1p mutants, and DTT-treated Ero1p (WT reduced). (A) The Trx red oxidation reaction was blocked with AMS and separated by SDS-PAGE under nonreducing conditions. The asterisk (*) indicates a species that migrates slower than Ero1p C90A/C349A when blocked with AMS, but faster than this disulfide mutant when blocked with NEM (compare with panel C). (B) The same experiment as in (A) was performed, but the samples were reduced with DTT after AMS treatment before separation by SDS-PAGE. (C) Reactions were blocked with NEM and applied to the gel under nonreducing conditions.

Techniques Used: Mutagenesis, Concentration Assay, Blocking Assay, Migration, SDS Page, Affinity Magnetic Separation

2) Product Images from "The Mitochondrial Intermembrane Space Oxireductase Mia40 Funnels the Oxidative Folding Pathway of the Cytochrome c Oxidase Assembly Protein Cox19 *"

Article Title: The Mitochondrial Intermembrane Space Oxireductase Mia40 Funnels the Oxidative Folding Pathway of the Cytochrome c Oxidase Assembly Protein Cox19 *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M114.553479

Reductive unfolding of Cox19 involves single disulfide intermediates. A , Cox19 reduction and unfolding can be monitored by Cox19 W74 fluorescence. RFU , relative fluorescence units. DTT 20 m m was added to oxidized Cox19 and W74 emission spectrum recorded
Figure Legend Snippet: Reductive unfolding of Cox19 involves single disulfide intermediates. A , Cox19 reduction and unfolding can be monitored by Cox19 W74 fluorescence. RFU , relative fluorescence units. DTT 20 m m was added to oxidized Cox19 and W74 emission spectrum recorded

Techniques Used: Fluorescence

Slow oxidation of Cox19 results in folding. A , Cox19 was reduced in 20 m m DTT for at least 3 h. The reduced protein was loaded in a small PD10 column containing a G25 resin. The recovered protein, free of DTT, was allowed to air oxidized at 25 °C,
Figure Legend Snippet: Slow oxidation of Cox19 results in folding. A , Cox19 was reduced in 20 m m DTT for at least 3 h. The reduced protein was loaded in a small PD10 column containing a G25 resin. The recovered protein, free of DTT, was allowed to air oxidized at 25 °C,

Techniques Used:

3) Product Images from "Crystal structure of peroxiredoxin 3 from Vibrio vulnificus and its implications for scavenging peroxides and nitric oxide"

Article Title: Crystal structure of peroxiredoxin 3 from Vibrio vulnificus and its implications for scavenging peroxides and nitric oxide

Journal: IUCrJ

doi: 10.1107/S205225251701750X

Intermolecular disulfide-bond formation in Vv Prx3 (C73S) on treatment with peroxide. Proteins were treated with H 2 O 2 ( a ) or tert -butyl hydrogen peroxide ( t -BOOH) ( b ) at the concentrations shown for 30 min and the reactions were stopped by adding iodoacetamide. Samples were boiled in the presence (+) or absence (−) of 10 m M DTT for SDS–PAGE analysis. ( c ) Determination of the second-order reaction rate constant for the reaction of Vv Prx3 and H 2 O 2 using competition kinetics with HRP. The fitting line was calculated by the least-squares method and the error bars reflect the standard deviation from three independent experiments. ( d ) Purified Vv Prx3 (C73S) (70 µ M ) was reacted with 50 µ M H 2 O 2 for the given times and analyzed by SDS–PAGE. At a time of 30 s, ∼30 µ M protein was linked by a disulfide bond as estimated using the density-measuring program ImageJ . The protein band for disulfide bond-containing Vv Prx3 (D) and bands that lack a disulfide bond (M) are indicated.
Figure Legend Snippet: Intermolecular disulfide-bond formation in Vv Prx3 (C73S) on treatment with peroxide. Proteins were treated with H 2 O 2 ( a ) or tert -butyl hydrogen peroxide ( t -BOOH) ( b ) at the concentrations shown for 30 min and the reactions were stopped by adding iodoacetamide. Samples were boiled in the presence (+) or absence (−) of 10 m M DTT for SDS–PAGE analysis. ( c ) Determination of the second-order reaction rate constant for the reaction of Vv Prx3 and H 2 O 2 using competition kinetics with HRP. The fitting line was calculated by the least-squares method and the error bars reflect the standard deviation from three independent experiments. ( d ) Purified Vv Prx3 (C73S) (70 µ M ) was reacted with 50 µ M H 2 O 2 for the given times and analyzed by SDS–PAGE. At a time of 30 s, ∼30 µ M protein was linked by a disulfide bond as estimated using the density-measuring program ImageJ . The protein band for disulfide bond-containing Vv Prx3 (D) and bands that lack a disulfide bond (M) are indicated.

Techniques Used: SDS Page, Standard Deviation, Purification

4) Product Images from "Blue Light-induced Dimerization of Monomeric Aureochrome-1 Enhances Its Affinity for the Target Sequence *"

Article Title: Blue Light-induced Dimerization of Monomeric Aureochrome-1 Enhances Its Affinity for the Target Sequence *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M114.554618

Elution profiles of monomeric ZLs with Apo. Elution profiles of ZLC 2 S ( A ), hZLC 2 S ( B and C ), and DTT-ZL ( D ) incubated with Apo in the presence of 300 m m ( A , C , and D ) or 250 m m ( B ) NaCl, in the dark ( upper ), light ( middle ), and light-dark ( lower ) states.
Figure Legend Snippet: Elution profiles of monomeric ZLs with Apo. Elution profiles of ZLC 2 S ( A ), hZLC 2 S ( B and C ), and DTT-ZL ( D ) incubated with Apo in the presence of 300 m m ( A , C , and D ) or 250 m m ( B ) NaCl, in the dark ( upper ), light ( middle ), and light-dark ( lower ) states.

Techniques Used: Incubation

Elution profiles of DTT-FL. A , elution profiles of DTT-FL in SEC buffer containing 250 m m NaCl and 5 m m DTT, in the dark ( solid lines ), light ( dashed lines ), and light-dark ( dotted lines ) states. Diamonds indicate elution volumes of the peaks in the dark
Figure Legend Snippet: Elution profiles of DTT-FL. A , elution profiles of DTT-FL in SEC buffer containing 250 m m NaCl and 5 m m DTT, in the dark ( solid lines ), light ( dashed lines ), and light-dark ( dotted lines ) states. Diamonds indicate elution volumes of the peaks in the dark

Techniques Used: Size-exclusion Chromatography

5) Product Images from "A preliminary crystallographic study of recombinant NicX, an Fe2+-dependent 2,5-dihydroxypyridine dioxygenase from Pseudomonas putida KT2440"

Article Title: A preliminary crystallographic study of recombinant NicX, an Fe2+-dependent 2,5-dihydroxypyridine dioxygenase from Pseudomonas putida KT2440

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

doi: 10.1107/S174430911001119X

Optimized crystals of recombinant NicX from P. putida KT2440 grown at 291 K in 22%( w / v ) PEG 4000, 0.1 M sodium acetate and 0.1 M HEPES pH 7.5 with 2 m M DTT. The final drops contained 1 µl protein solution
Figure Legend Snippet: Optimized crystals of recombinant NicX from P. putida KT2440 grown at 291 K in 22%( w / v ) PEG 4000, 0.1 M sodium acetate and 0.1 M HEPES pH 7.5 with 2 m M DTT. The final drops contained 1 µl protein solution

Techniques Used: Recombinant

Related Articles

Size-exclusion Chromatography:

Article Title: Structural and Functional Characterization of the Recombinant Death Domain from Death-Associated Protein Kinase
Article Snippet: .. The gel bands were stained with Coomassie blue dye. ( C ) Affinity and SEC purified GB1-FADD-DD in 20 mM NaPi, 150 mM NaCl, 3 mM DTT, pH 6.2 and GB1-DAPk-DD(S) in 20 mM NaPi, 150 mM NaCl, 3 mM DTT, pH 7.4 were applied on a pre-packed Superose-12 analytical size-exclusion column (Amersham Biosciences) at the same concentration, i.e., 1 mg/ml. .. At a lower concentration, the purified GB1-DAPk-DD(S) protein elutes at an elution volume near identical to the GB1-FADD-DD protein, which does not form oligomers.

Purification:

Article Title: Structural and Functional Characterization of the Recombinant Death Domain from Death-Associated Protein Kinase
Article Snippet: .. The gel bands were stained with Coomassie blue dye. ( C ) Affinity and SEC purified GB1-FADD-DD in 20 mM NaPi, 150 mM NaCl, 3 mM DTT, pH 6.2 and GB1-DAPk-DD(S) in 20 mM NaPi, 150 mM NaCl, 3 mM DTT, pH 7.4 were applied on a pre-packed Superose-12 analytical size-exclusion column (Amersham Biosciences) at the same concentration, i.e., 1 mg/ml. .. At a lower concentration, the purified GB1-DAPk-DD(S) protein elutes at an elution volume near identical to the GB1-FADD-DD protein, which does not form oligomers.

Concentration Assay:

Article Title: Structural and Functional Characterization of the Recombinant Death Domain from Death-Associated Protein Kinase
Article Snippet: .. The gel bands were stained with Coomassie blue dye. ( C ) Affinity and SEC purified GB1-FADD-DD in 20 mM NaPi, 150 mM NaCl, 3 mM DTT, pH 6.2 and GB1-DAPk-DD(S) in 20 mM NaPi, 150 mM NaCl, 3 mM DTT, pH 7.4 were applied on a pre-packed Superose-12 analytical size-exclusion column (Amersham Biosciences) at the same concentration, i.e., 1 mg/ml. .. At a lower concentration, the purified GB1-DAPk-DD(S) protein elutes at an elution volume near identical to the GB1-FADD-DD protein, which does not form oligomers.

Incubation:

Article Title: The low-affinity complex of cytochrome c and its peroxidase
Article Snippet: .. First, to reduce the possible intermolecular disulfides, a relevant single-cysteine CcP variant was incubated with a 10-fold molar excess of DTT for 1 h at room temperature (RT) and then exchanged into 20 mM Tris-HCl 100 mM NaCl pH 8.0 on a HiTrap Desalting column (GE Healthcare), concomitantly removing the reductant. .. Second, the CcP—now bearing a free thiol group—was incubated with a 10-fold molar excess of 5,5′-dithiobis-(2-nitrobenzoate) [DTNB or Ellman's reagent] for 1 h at RT, yielding the modified protein and the yellow-coloured TNB.

Article Title: Characterization of the ?? subcomplex of Pseudomonas aeruginosa DNA polymerase III
Article Snippet: .. After washing with GS buffer (50 mM Tris-HCl pH 7.0, 150 mM NaCl, 10% glycerol (v/v), 1 mM EDTA, 1 mM DTT), the column was incubated overnight at 4°C with 840 μg of PreScission™ protease (GE Healthcare Life Sciences). ..

Staining:

Article Title: Structural and Functional Characterization of the Recombinant Death Domain from Death-Associated Protein Kinase
Article Snippet: .. The gel bands were stained with Coomassie blue dye. ( C ) Affinity and SEC purified GB1-FADD-DD in 20 mM NaPi, 150 mM NaCl, 3 mM DTT, pH 6.2 and GB1-DAPk-DD(S) in 20 mM NaPi, 150 mM NaCl, 3 mM DTT, pH 7.4 were applied on a pre-packed Superose-12 analytical size-exclusion column (Amersham Biosciences) at the same concentration, i.e., 1 mg/ml. .. At a lower concentration, the purified GB1-DAPk-DD(S) protein elutes at an elution volume near identical to the GB1-FADD-DD protein, which does not form oligomers.

Variant Assay:

Article Title: The low-affinity complex of cytochrome c and its peroxidase
Article Snippet: .. First, to reduce the possible intermolecular disulfides, a relevant single-cysteine CcP variant was incubated with a 10-fold molar excess of DTT for 1 h at room temperature (RT) and then exchanged into 20 mM Tris-HCl 100 mM NaCl pH 8.0 on a HiTrap Desalting column (GE Healthcare), concomitantly removing the reductant. .. Second, the CcP—now bearing a free thiol group—was incubated with a 10-fold molar excess of 5,5′-dithiobis-(2-nitrobenzoate) [DTNB or Ellman's reagent] for 1 h at RT, yielding the modified protein and the yellow-coloured TNB.

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    GE Healthcare m dtt
    Reductive unfolding of <t>Cox19</t> involves single disulfide intermediates. A , Cox19 reduction and unfolding can be monitored by Cox19 W74 fluorescence. RFU , relative fluorescence units. <t>DTT</t> 20 m m was added to oxidized Cox19 and W74 emission spectrum recorded
    M Dtt, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m dtt/product/GE Healthcare
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    m dtt - by Bioz Stars, 2020-09
    99/100 stars
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    Reductive unfolding of Cox19 involves single disulfide intermediates. A , Cox19 reduction and unfolding can be monitored by Cox19 W74 fluorescence. RFU , relative fluorescence units. DTT 20 m m was added to oxidized Cox19 and W74 emission spectrum recorded

    Journal: The Journal of Biological Chemistry

    Article Title: The Mitochondrial Intermembrane Space Oxireductase Mia40 Funnels the Oxidative Folding Pathway of the Cytochrome c Oxidase Assembly Protein Cox19 *

    doi: 10.1074/jbc.M114.553479

    Figure Lengend Snippet: Reductive unfolding of Cox19 involves single disulfide intermediates. A , Cox19 reduction and unfolding can be monitored by Cox19 W74 fluorescence. RFU , relative fluorescence units. DTT 20 m m was added to oxidized Cox19 and W74 emission spectrum recorded

    Article Snippet: For oxidative folding, Cox19, previously reduced with 20 m m DTT was loaded in a PD mini trap G-25 column (GE) containing Sephadex G-25, and the protein free of reducing agent was recovered in Tris 50 m m , pH 8.4 NaCl 100 m m following the manufacturer's instructions.

    Techniques: Fluorescence

    Slow oxidation of Cox19 results in folding. A , Cox19 was reduced in 20 m m DTT for at least 3 h. The reduced protein was loaded in a small PD10 column containing a G25 resin. The recovered protein, free of DTT, was allowed to air oxidized at 25 °C,

    Journal: The Journal of Biological Chemistry

    Article Title: The Mitochondrial Intermembrane Space Oxireductase Mia40 Funnels the Oxidative Folding Pathway of the Cytochrome c Oxidase Assembly Protein Cox19 *

    doi: 10.1074/jbc.M114.553479

    Figure Lengend Snippet: Slow oxidation of Cox19 results in folding. A , Cox19 was reduced in 20 m m DTT for at least 3 h. The reduced protein was loaded in a small PD10 column containing a G25 resin. The recovered protein, free of DTT, was allowed to air oxidized at 25 °C,

    Article Snippet: For oxidative folding, Cox19, previously reduced with 20 m m DTT was loaded in a PD mini trap G-25 column (GE) containing Sephadex G-25, and the protein free of reducing agent was recovered in Tris 50 m m , pH 8.4 NaCl 100 m m following the manufacturer's instructions.

    Techniques:

    Disulfide reduction during catalysis by Ero1p and the de-regulated mutant C150A/C295A. Enzyme at a final concentration of 2 μ M was mixed with 100 μ M Trx red . Aliquots were removed from the reactions at the indicated time points, and disulfide status was preserved by blocking with maleimide reagents. The migration of Ero1p on SDS-PAGE at the various time points was examined and compared with wild-type Ero1p (WT), the indicated Ero1p mutants, and DTT-treated Ero1p (WT reduced). (A) The Trx red oxidation reaction was blocked with AMS and separated by SDS-PAGE under nonreducing conditions. The asterisk (*) indicates a species that migrates slower than Ero1p C90A/C349A when blocked with AMS, but faster than this disulfide mutant when blocked with NEM (compare with panel C). (B) The same experiment as in (A) was performed, but the samples were reduced with DTT after AMS treatment before separation by SDS-PAGE. (C) Reactions were blocked with NEM and applied to the gel under nonreducing conditions.

    Journal: Protein Science : A Publication of the Protein Society

    Article Title: Steps in reductive activation of the disulfide-generating enzyme Ero1p

    doi: 10.1002/pro.473

    Figure Lengend Snippet: Disulfide reduction during catalysis by Ero1p and the de-regulated mutant C150A/C295A. Enzyme at a final concentration of 2 μ M was mixed with 100 μ M Trx red . Aliquots were removed from the reactions at the indicated time points, and disulfide status was preserved by blocking with maleimide reagents. The migration of Ero1p on SDS-PAGE at the various time points was examined and compared with wild-type Ero1p (WT), the indicated Ero1p mutants, and DTT-treated Ero1p (WT reduced). (A) The Trx red oxidation reaction was blocked with AMS and separated by SDS-PAGE under nonreducing conditions. The asterisk (*) indicates a species that migrates slower than Ero1p C90A/C349A when blocked with AMS, but faster than this disulfide mutant when blocked with NEM (compare with panel C). (B) The same experiment as in (A) was performed, but the samples were reduced with DTT after AMS treatment before separation by SDS-PAGE. (C) Reactions were blocked with NEM and applied to the gel under nonreducing conditions.

    Article Snippet: For oxidation assays, Trx was reduced with 100 m M DTT for 1 h and then desalted using a PD-10 column (GE Healthcare) equilibrated in 50 m M phosphate buffer, pH 7.5, 65 m M NaCl, 0.5 m M EDTA.

    Techniques: Mutagenesis, Concentration Assay, Blocking Assay, Migration, SDS Page, Affinity Magnetic Separation

    Intermolecular disulfide-bond formation in Vv Prx3 (C73S) on treatment with peroxide. Proteins were treated with H 2 O 2 ( a ) or tert -butyl hydrogen peroxide ( t -BOOH) ( b ) at the concentrations shown for 30 min and the reactions were stopped by adding iodoacetamide. Samples were boiled in the presence (+) or absence (−) of 10 m M DTT for SDS–PAGE analysis. ( c ) Determination of the second-order reaction rate constant for the reaction of Vv Prx3 and H 2 O 2 using competition kinetics with HRP. The fitting line was calculated by the least-squares method and the error bars reflect the standard deviation from three independent experiments. ( d ) Purified Vv Prx3 (C73S) (70 µ M ) was reacted with 50 µ M H 2 O 2 for the given times and analyzed by SDS–PAGE. At a time of 30 s, ∼30 µ M protein was linked by a disulfide bond as estimated using the density-measuring program ImageJ . The protein band for disulfide bond-containing Vv Prx3 (D) and bands that lack a disulfide bond (M) are indicated.

    Journal: IUCrJ

    Article Title: Crystal structure of peroxiredoxin 3 from Vibrio vulnificus and its implications for scavenging peroxides and nitric oxide

    doi: 10.1107/S205225251701750X

    Figure Lengend Snippet: Intermolecular disulfide-bond formation in Vv Prx3 (C73S) on treatment with peroxide. Proteins were treated with H 2 O 2 ( a ) or tert -butyl hydrogen peroxide ( t -BOOH) ( b ) at the concentrations shown for 30 min and the reactions were stopped by adding iodoacetamide. Samples were boiled in the presence (+) or absence (−) of 10 m M DTT for SDS–PAGE analysis. ( c ) Determination of the second-order reaction rate constant for the reaction of Vv Prx3 and H 2 O 2 using competition kinetics with HRP. The fitting line was calculated by the least-squares method and the error bars reflect the standard deviation from three independent experiments. ( d ) Purified Vv Prx3 (C73S) (70 µ M ) was reacted with 50 µ M H 2 O 2 for the given times and analyzed by SDS–PAGE. At a time of 30 s, ∼30 µ M protein was linked by a disulfide bond as estimated using the density-measuring program ImageJ . The protein band for disulfide bond-containing Vv Prx3 (D) and bands that lack a disulfide bond (M) are indicated.

    Article Snippet: To prepare the reduced Vv Prx3 (C73S) protein, 10 m M DTT was added to the purified Vv Prx3 (C73S) protein and was then removed using a HiPrep 26/10 Desalting column (GE Healthcare, USA) pre-equilibrated with 20 m M Tris–HCl pH 8.0 buffer containing 150 m M NaCl.

    Techniques: SDS Page, Standard Deviation, Purification

    Elution profiles of monomeric ZLs with Apo. Elution profiles of ZLC 2 S ( A ), hZLC 2 S ( B and C ), and DTT-ZL ( D ) incubated with Apo in the presence of 300 m m ( A , C , and D ) or 250 m m ( B ) NaCl, in the dark ( upper ), light ( middle ), and light-dark ( lower ) states.

    Journal: The Journal of Biological Chemistry

    Article Title: Blue Light-induced Dimerization of Monomeric Aureochrome-1 Enhances Its Affinity for the Target Sequence *

    doi: 10.1074/jbc.M114.554618

    Figure Lengend Snippet: Elution profiles of monomeric ZLs with Apo. Elution profiles of ZLC 2 S ( A ), hZLC 2 S ( B and C ), and DTT-ZL ( D ) incubated with Apo in the presence of 300 m m ( A , C , and D ) or 250 m m ( B ) NaCl, in the dark ( upper ), light ( middle ), and light-dark ( lower ) states.

    Article Snippet: After exchanging the solvents to the loading buffer (400 m m NaCl, 20 m m Tris-HCl, pH 7.0) containing 2 m m DTT using a gel filtration column (PD-10, GE Healthcare), the recombinant proteins were stored at 4 °C until use.

    Techniques: Incubation

    Elution profiles of DTT-FL. A , elution profiles of DTT-FL in SEC buffer containing 250 m m NaCl and 5 m m DTT, in the dark ( solid lines ), light ( dashed lines ), and light-dark ( dotted lines ) states. Diamonds indicate elution volumes of the peaks in the dark

    Journal: The Journal of Biological Chemistry

    Article Title: Blue Light-induced Dimerization of Monomeric Aureochrome-1 Enhances Its Affinity for the Target Sequence *

    doi: 10.1074/jbc.M114.554618

    Figure Lengend Snippet: Elution profiles of DTT-FL. A , elution profiles of DTT-FL in SEC buffer containing 250 m m NaCl and 5 m m DTT, in the dark ( solid lines ), light ( dashed lines ), and light-dark ( dotted lines ) states. Diamonds indicate elution volumes of the peaks in the dark

    Article Snippet: After exchanging the solvents to the loading buffer (400 m m NaCl, 20 m m Tris-HCl, pH 7.0) containing 2 m m DTT using a gel filtration column (PD-10, GE Healthcare), the recombinant proteins were stored at 4 °C until use.

    Techniques: Size-exclusion Chromatography