bistris protein gels  (Thermo Fisher)


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    Name:
    NuPAGE 12 Bis Tris Protein Gels
    Description:
    Invitrogen NuPAGE Bis Tris protein gels are precast polyacrylamide gels designed to give optimal separation of a wide range of proteins under denaturing conditions Unlike traditional Tris glycine gels NuPAGE Bis Tris gels have a neutral pH environment that minimizes protein modifications Use NuPAGE Bis Tris gels for preparing proteins for sequencing mass spectrometry and any other techniques where protein integrity is crucial Also use NuPAGE gels for optimal results during day to day use Features of NuPAGE Bis Tris gels • Better protein integrity optimized sample preparation process preserves your proteins • Wide ranges of molecular weight separation select the right gel and running buffer to get the optimal separation of your proteins • Faster run times get separation of your proteins in as little as 35 minutes • Longer shelf life NuPAGE Bis Tris gels can be stored for at least 12 months at room temperature Learn more about all of our NuPAGE Bis Tris gels View migration charts › Choose the right NuPAGE Bis Tris gel for your protein separation Obtain optimal separation of your proteins by choosing the right combination of gel and running buffer NuPAGE Bis Tris protein gels come in four polyacrylamide concentrations 8 10 12 and a 4 12 gradient Gels come in two sizes mini 8 cm x 8 cm or midi 8 7 cm x 13 3 cm and either 1 0 mm mini and midi gels or 1 5 mm mini gel format only in thickness NuPAGE Bis Tris gels also come in multiple well formats NuPAGE Bis Tris gels are formulated for denaturing gel electrophoresis applications For optimal sample preparation use the NuPAGE LDS Sample Buffer and NuPAGE Sample Reducing Agent Use NuPAGE Antioxidant in the running buffer to maintain the reduced state of the proteins during the run and to allow maximum band sharpness The gels can be run using NuPAGE MES SDS Running Buffer to better resolve small proteins or NuPAGE MOPS SDS Running Buffer to resolve medium to large size proteins We also provide NuPAGE Tris Acetate gels for separating larger proteins For classic Laemmli based Tris glycine electrophoresis we provide Novex Tris Glycine gels For transfer of proteins to a membrane we recommend using NuPAGE Transfer Buffer Rapid semi dry transfer can be done using the Pierce Power Blotter or rapid dry transfer using the iBlot 2 Gel Transfer Device Alternatively traditional wet transfer can be performed using the XCell II Blot Module or the Mini Blot Module Related links Overview of 1D Protein Electrophoresis Comparison of NuPAGE Tris Bis vs traditional Tris glycine gels
    Catalog Number:
    np0341box
    Price:
    None
    Applications:
    1D Gel Electrophoresis|NuPAGE® Novex® Bis-Tris Gel Electrophoresis|Protein Biology|Protein Gel Electrophoresis
    Category:
    Gels Fractionation Strips Membranes
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    Structured Review

    Thermo Fisher bistris protein gels
    Inhibition of the auto-catalytic activation of FhCL1 by rFhKT1 and rFhKT1Leu 15 /Arg 15 . M, molecular mass marker; lane 1, FhCL1 zymogen; lane 2, mature FhCL1 following auto-catalytic activation at pH 4.5 for 1 h; lane 3, FhCL1 and rFhKT1 following incubation at pH 4.5 for 1 h; lane 4, FhCL1 and rFhKT1Leu 15 /Arg 15 following incubation at pH 4.5 for 1 h; lane 5, FhCL1 and bovine pancreatic trypsin inhibitor following incubation at pH 4.5 for 1 h. Double white arrowhead indicates FhCL1 zymogen; double gray arrowhead indicates mature FhCL1, and double black arrowhead indicates FhCL1 pro-peptide removed following auto-catalytic activation. Single white arrowhead indicates position of FhKT1; single black arrowhead indicates FhKT1Leu 15 /Arg 15 , and single gray arrowhead shows position of BPTI. Proteins were separated on a NuPAGE Novex 4–12% <t>BisTris</t> protein gel and stained with Biosafe Coomassie (Bio-Rad). M, molecular mass markers.
    Invitrogen NuPAGE Bis Tris protein gels are precast polyacrylamide gels designed to give optimal separation of a wide range of proteins under denaturing conditions Unlike traditional Tris glycine gels NuPAGE Bis Tris gels have a neutral pH environment that minimizes protein modifications Use NuPAGE Bis Tris gels for preparing proteins for sequencing mass spectrometry and any other techniques where protein integrity is crucial Also use NuPAGE gels for optimal results during day to day use Features of NuPAGE Bis Tris gels • Better protein integrity optimized sample preparation process preserves your proteins • Wide ranges of molecular weight separation select the right gel and running buffer to get the optimal separation of your proteins • Faster run times get separation of your proteins in as little as 35 minutes • Longer shelf life NuPAGE Bis Tris gels can be stored for at least 12 months at room temperature Learn more about all of our NuPAGE Bis Tris gels View migration charts › Choose the right NuPAGE Bis Tris gel for your protein separation Obtain optimal separation of your proteins by choosing the right combination of gel and running buffer NuPAGE Bis Tris protein gels come in four polyacrylamide concentrations 8 10 12 and a 4 12 gradient Gels come in two sizes mini 8 cm x 8 cm or midi 8 7 cm x 13 3 cm and either 1 0 mm mini and midi gels or 1 5 mm mini gel format only in thickness NuPAGE Bis Tris gels also come in multiple well formats NuPAGE Bis Tris gels are formulated for denaturing gel electrophoresis applications For optimal sample preparation use the NuPAGE LDS Sample Buffer and NuPAGE Sample Reducing Agent Use NuPAGE Antioxidant in the running buffer to maintain the reduced state of the proteins during the run and to allow maximum band sharpness The gels can be run using NuPAGE MES SDS Running Buffer to better resolve small proteins or NuPAGE MOPS SDS Running Buffer to resolve medium to large size proteins We also provide NuPAGE Tris Acetate gels for separating larger proteins For classic Laemmli based Tris glycine electrophoresis we provide Novex Tris Glycine gels For transfer of proteins to a membrane we recommend using NuPAGE Transfer Buffer Rapid semi dry transfer can be done using the Pierce Power Blotter or rapid dry transfer using the iBlot 2 Gel Transfer Device Alternatively traditional wet transfer can be performed using the XCell II Blot Module or the Mini Blot Module Related links Overview of 1D Protein Electrophoresis Comparison of NuPAGE Tris Bis vs traditional Tris glycine gels
    https://www.bioz.com/result/bistris protein gels/product/Thermo Fisher
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    bistris protein gels - by Bioz Stars, 2020-02
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    Images

    1) Product Images from "Unexpected Activity of a Novel Kunitz-type Inhibitor"

    Article Title: Unexpected Activity of a Novel Kunitz-type Inhibitor

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M116.724344

    Inhibition of the auto-catalytic activation of FhCL1 by rFhKT1 and rFhKT1Leu 15 /Arg 15 . M, molecular mass marker; lane 1, FhCL1 zymogen; lane 2, mature FhCL1 following auto-catalytic activation at pH 4.5 for 1 h; lane 3, FhCL1 and rFhKT1 following incubation at pH 4.5 for 1 h; lane 4, FhCL1 and rFhKT1Leu 15 /Arg 15 following incubation at pH 4.5 for 1 h; lane 5, FhCL1 and bovine pancreatic trypsin inhibitor following incubation at pH 4.5 for 1 h. Double white arrowhead indicates FhCL1 zymogen; double gray arrowhead indicates mature FhCL1, and double black arrowhead indicates FhCL1 pro-peptide removed following auto-catalytic activation. Single white arrowhead indicates position of FhKT1; single black arrowhead indicates FhKT1Leu 15 /Arg 15 , and single gray arrowhead shows position of BPTI. Proteins were separated on a NuPAGE Novex 4–12% BisTris protein gel and stained with Biosafe Coomassie (Bio-Rad). M, molecular mass markers.
    Figure Legend Snippet: Inhibition of the auto-catalytic activation of FhCL1 by rFhKT1 and rFhKT1Leu 15 /Arg 15 . M, molecular mass marker; lane 1, FhCL1 zymogen; lane 2, mature FhCL1 following auto-catalytic activation at pH 4.5 for 1 h; lane 3, FhCL1 and rFhKT1 following incubation at pH 4.5 for 1 h; lane 4, FhCL1 and rFhKT1Leu 15 /Arg 15 following incubation at pH 4.5 for 1 h; lane 5, FhCL1 and bovine pancreatic trypsin inhibitor following incubation at pH 4.5 for 1 h. Double white arrowhead indicates FhCL1 zymogen; double gray arrowhead indicates mature FhCL1, and double black arrowhead indicates FhCL1 pro-peptide removed following auto-catalytic activation. Single white arrowhead indicates position of FhKT1; single black arrowhead indicates FhKT1Leu 15 /Arg 15 , and single gray arrowhead shows position of BPTI. Proteins were separated on a NuPAGE Novex 4–12% BisTris protein gel and stained with Biosafe Coomassie (Bio-Rad). M, molecular mass markers.

    Techniques Used: Inhibition, Activation Assay, Marker, Incubation, Staining

    Structural representation of FhKT1 and FhKT1Leu 15 /Arg 15 and their recombinant expression. A, sequence alignment of BPTI, FhKT1, and FhKT1Leu 15 /Arg 15 . The asterisk denotes the P1 site at position 15. Lines indicate the conserved disulfide bonds that occur between Cys 1 and Cys 6 , Cys 2 and Cys 4 , and Cys 3 and Cys 5 , with cysteine residues highlighted in black bars. B, recombinant forms of FhKT1 and FhKT1Leu 15 /Arg 15 were expressed as secretory proteins in the methylotrophic yeast P. pastoris with a yield of ∼5–10 mg of soluble protein from each 1 liter of culture. rFhKT1 ( lane 1 ) and rFhKT1Leu 15 /Arg 15 ( lane 2 ) were isolated by NTA-affinity chromatography and analyzed by NuPAGE Novex 4–12% BisTris protein gel, stained with Biosafe Coomassie (Bio-Rad). C, homology model of FhKT1 built based on BPTI (PDB code 3OTJ ) displaying the three disulfide bonds ( yellow ), the α-helix ( red ), and anti-parallel β-sheets ( blue ) characteristic of KT protease inhibitors. The P1 Leu 15 located at the peak of the reactive loop is also shown in gray .
    Figure Legend Snippet: Structural representation of FhKT1 and FhKT1Leu 15 /Arg 15 and their recombinant expression. A, sequence alignment of BPTI, FhKT1, and FhKT1Leu 15 /Arg 15 . The asterisk denotes the P1 site at position 15. Lines indicate the conserved disulfide bonds that occur between Cys 1 and Cys 6 , Cys 2 and Cys 4 , and Cys 3 and Cys 5 , with cysteine residues highlighted in black bars. B, recombinant forms of FhKT1 and FhKT1Leu 15 /Arg 15 were expressed as secretory proteins in the methylotrophic yeast P. pastoris with a yield of ∼5–10 mg of soluble protein from each 1 liter of culture. rFhKT1 ( lane 1 ) and rFhKT1Leu 15 /Arg 15 ( lane 2 ) were isolated by NTA-affinity chromatography and analyzed by NuPAGE Novex 4–12% BisTris protein gel, stained with Biosafe Coomassie (Bio-Rad). C, homology model of FhKT1 built based on BPTI (PDB code 3OTJ ) displaying the three disulfide bonds ( yellow ), the α-helix ( red ), and anti-parallel β-sheets ( blue ) characteristic of KT protease inhibitors. The P1 Leu 15 located at the peak of the reactive loop is also shown in gray .

    Techniques Used: Recombinant, Expressing, Sequencing, Isolation, Affinity Chromatography, Staining

    2) Product Images from "Unexpected Activity of a Novel Kunitz-type Inhibitor"

    Article Title: Unexpected Activity of a Novel Kunitz-type Inhibitor

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M116.724344

    Inhibition of the auto-catalytic activation of FhCL1 by rFhKT1 and rFhKT1Leu 15 /Arg 15 . M, molecular mass marker; lane 1, FhCL1 zymogen; lane 2, mature FhCL1 following auto-catalytic activation at pH 4.5 for 1 h; lane 3, FhCL1 and rFhKT1 following incubation at pH 4.5 for 1 h; lane 4, FhCL1 and rFhKT1Leu 15 /Arg 15 following incubation at pH 4.5 for 1 h; lane 5, FhCL1 and bovine pancreatic trypsin inhibitor following incubation at pH 4.5 for 1 h. Double white arrowhead indicates FhCL1 zymogen; double gray arrowhead indicates mature FhCL1, and double black arrowhead indicates FhCL1 pro-peptide removed following auto-catalytic activation. Single white arrowhead indicates position of FhKT1; single black arrowhead indicates FhKT1Leu 15 /Arg 15 , and single gray arrowhead shows position of BPTI. Proteins were separated on a NuPAGE Novex 4–12% BisTris protein gel and stained with Biosafe Coomassie (Bio-Rad). M, molecular mass markers.
    Figure Legend Snippet: Inhibition of the auto-catalytic activation of FhCL1 by rFhKT1 and rFhKT1Leu 15 /Arg 15 . M, molecular mass marker; lane 1, FhCL1 zymogen; lane 2, mature FhCL1 following auto-catalytic activation at pH 4.5 for 1 h; lane 3, FhCL1 and rFhKT1 following incubation at pH 4.5 for 1 h; lane 4, FhCL1 and rFhKT1Leu 15 /Arg 15 following incubation at pH 4.5 for 1 h; lane 5, FhCL1 and bovine pancreatic trypsin inhibitor following incubation at pH 4.5 for 1 h. Double white arrowhead indicates FhCL1 zymogen; double gray arrowhead indicates mature FhCL1, and double black arrowhead indicates FhCL1 pro-peptide removed following auto-catalytic activation. Single white arrowhead indicates position of FhKT1; single black arrowhead indicates FhKT1Leu 15 /Arg 15 , and single gray arrowhead shows position of BPTI. Proteins were separated on a NuPAGE Novex 4–12% BisTris protein gel and stained with Biosafe Coomassie (Bio-Rad). M, molecular mass markers.

    Techniques Used: Inhibition, Activation Assay, Marker, Incubation, Staining

    Structural representation of FhKT1 and FhKT1Leu 15 /Arg 15 and their recombinant expression. A, sequence alignment of BPTI, FhKT1, and FhKT1Leu 15 /Arg 15 . The asterisk denotes the P1 site at position 15. Lines indicate the conserved disulfide bonds that occur between Cys 1 and Cys 6 , Cys 2 and Cys 4 , and Cys 3 and Cys 5 , with cysteine residues highlighted in black bars. B, recombinant forms of FhKT1 and FhKT1Leu 15 /Arg 15 were expressed as secretory proteins in the methylotrophic yeast P. pastoris with a yield of ∼5–10 mg of soluble protein from each 1 liter of culture. rFhKT1 ( lane 1 ) and rFhKT1Leu 15 /Arg 15 ( lane 2 ) were isolated by NTA-affinity chromatography and analyzed by NuPAGE Novex 4–12% BisTris protein gel, stained with Biosafe Coomassie (Bio-Rad). C, homology model of FhKT1 built based on BPTI (PDB code 3OTJ ) displaying the three disulfide bonds ( yellow ), the α-helix ( red ), and anti-parallel β-sheets ( blue ) characteristic of KT protease inhibitors. The P1 Leu 15 located at the peak of the reactive loop is also shown in gray .
    Figure Legend Snippet: Structural representation of FhKT1 and FhKT1Leu 15 /Arg 15 and their recombinant expression. A, sequence alignment of BPTI, FhKT1, and FhKT1Leu 15 /Arg 15 . The asterisk denotes the P1 site at position 15. Lines indicate the conserved disulfide bonds that occur between Cys 1 and Cys 6 , Cys 2 and Cys 4 , and Cys 3 and Cys 5 , with cysteine residues highlighted in black bars. B, recombinant forms of FhKT1 and FhKT1Leu 15 /Arg 15 were expressed as secretory proteins in the methylotrophic yeast P. pastoris with a yield of ∼5–10 mg of soluble protein from each 1 liter of culture. rFhKT1 ( lane 1 ) and rFhKT1Leu 15 /Arg 15 ( lane 2 ) were isolated by NTA-affinity chromatography and analyzed by NuPAGE Novex 4–12% BisTris protein gel, stained with Biosafe Coomassie (Bio-Rad). C, homology model of FhKT1 built based on BPTI (PDB code 3OTJ ) displaying the three disulfide bonds ( yellow ), the α-helix ( red ), and anti-parallel β-sheets ( blue ) characteristic of KT protease inhibitors. The P1 Leu 15 located at the peak of the reactive loop is also shown in gray .

    Techniques Used: Recombinant, Expressing, Sequencing, Isolation, Affinity Chromatography, Staining

    3) Product Images from "IRE1–XBP1 pathway regulates oxidative proinsulin folding in pancreatic β cells"

    Article Title: IRE1–XBP1 pathway regulates oxidative proinsulin folding in pancreatic β cells

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201707143

    Detection of mixed-disulfide complexes between proinsulin and PDI family members in MIN6 cells and reconstitution of the five PDIs in MIN6 ( Ire1α ΔR/ΔR ) cells. (A) Cys-mediated interaction between PDI family proteins and newly synthesized proinsulin during proinsulin folding. WT MIN6 cells were transiently transfected with indicated FLAG-tagged PDI family proteins, pulsed for 5 min with [ 35 S]Met/Cys, directly treated with TCA, and subjected to alkylation with NEM. Mixed-disulfide complexes between PDIs and proinsulin were collected by consecutive immunoprecipitation (first immunoprecipitation by anti-FLAG antibody; second immunoprecipitation by antiproinsulin antibody). After elution, samples were treated with (Reducing) or without (Nonreducing) DTT, separated by NuPAGE, and detected by autoradiography. (B) MIN6 ( Ire1α ΔR/ΔR ) cells were transiently transfected with WT five PDIs or five CM PDIs. The expression of insulin was induced by culturing cells for 4 h in the presence of low glucose (LG; L; 1.67 mM glucose/KRBH; 4 h) or high glucose (HG; H; 16.7 mM glucose/KRBH; 4 h), and the amounts of secreted insulin were measured by ELISA. White bars indicate insulin secretion upon exposure to low glucose. Black bars indicate insulin secretion upon exposure to high glucose. (C) MIN6 ( Ire1α fl/fl ) cells were transiently transfected with WT PDI/PDIA1 or WT five PDIs. The expression of insulin was induced, and amounts of secreted insulin were measured as described in B. n = 3. **, P
    Figure Legend Snippet: Detection of mixed-disulfide complexes between proinsulin and PDI family members in MIN6 cells and reconstitution of the five PDIs in MIN6 ( Ire1α ΔR/ΔR ) cells. (A) Cys-mediated interaction between PDI family proteins and newly synthesized proinsulin during proinsulin folding. WT MIN6 cells were transiently transfected with indicated FLAG-tagged PDI family proteins, pulsed for 5 min with [ 35 S]Met/Cys, directly treated with TCA, and subjected to alkylation with NEM. Mixed-disulfide complexes between PDIs and proinsulin were collected by consecutive immunoprecipitation (first immunoprecipitation by anti-FLAG antibody; second immunoprecipitation by antiproinsulin antibody). After elution, samples were treated with (Reducing) or without (Nonreducing) DTT, separated by NuPAGE, and detected by autoradiography. (B) MIN6 ( Ire1α ΔR/ΔR ) cells were transiently transfected with WT five PDIs or five CM PDIs. The expression of insulin was induced by culturing cells for 4 h in the presence of low glucose (LG; L; 1.67 mM glucose/KRBH; 4 h) or high glucose (HG; H; 16.7 mM glucose/KRBH; 4 h), and the amounts of secreted insulin were measured by ELISA. White bars indicate insulin secretion upon exposure to low glucose. Black bars indicate insulin secretion upon exposure to high glucose. (C) MIN6 ( Ire1α fl/fl ) cells were transiently transfected with WT PDI/PDIA1 or WT five PDIs. The expression of insulin was induced, and amounts of secreted insulin were measured as described in B. n = 3. **, P

    Techniques Used: Synthesized, Transfection, Immunoprecipitation, Autoradiography, Expressing, Enzyme-linked Immunosorbent Assay

    Decreased oxidative folding of proinsulin in IRE1α-RNase domain KO MIN6 cells. (A) Levels of Ins genes and transcription factors that regulate Ins genes relative to Gapdh expression levels in the indicated MIN6 cells were measured by qRT-PCR. White bars indicate control MIN6 ( Ire1α fl/fl ) cells (Con). Blue bars indicate MIN6 ( Ire1α fl/fl ) cells infected with Ad-GFP. Red bars indicate MIN6 ( Ire1α ΔR/ΔR ) cells infected with Ad-Cre. (B) To detect newly synthesized preproinsulin and proinsulin in the indicated MIN6 cells, MIN6 cells were pulsed with [ 35 S]Met/Cys for 30 min. Radiolabeled preproinsulin and proinsulin were then immunoprecipitated, separated by NuPAGE, and detected by autoradiography. The ratio of the translation of preproinsulin and proinsulin in MIN6 ( Ire1α ΔR/ΔR ) cells to that of MIN6 ( Ire1α fl/fl ) cells was quantified. Blue bar, MIN6 ( Ire1α fl/fl ) cells; red bar, MIN6 ( Ire1α ΔR/ΔR ) cells. (C) To compare the rate of conversion from preproinsulin to proinsulin in MIN6 ( Ire1α fl/fl or Ire1α ΔR/ΔR ) cells, the cells were pulsed for 1 min with [ 35 S]Met/Cys and then chased for the indicated times. The ratio of proinsulin to the sum of preproinsulin and proinsulin was quantified. The values for MIN6 ( Ire1α fl/fl ) and MIN6 ( Ire1α ΔR/ΔR ) cells are indicated by a blue solid line and by a red broken line, respectively. The values obtained did not differ significantly among time points by Student’s t test. (D) To compare the rate of oxidative folding of proinsulin in MIN6 ( Ire1α fl/fl or Ire1α ΔR/ΔR ) cells, the cells were pulsed for 5 min with [ 35 S]Met/Cys and then chased for the indicated times. Radiolabeled proinsulin was immunoprecipitated with an antiproinsulin antibody, boiled with (Reducing) or without (Nonreducing) DTT, separated by NuPAGE, and detected by autoradiography. The closed diamond indicates preproinsulin. (E) The ratio of oxidized proinsulin (left, red arrowhead) to total proinsulin (right, white arrowhead) in D was quantified. Error bars represent means and SD for each group. n = 3. *, P
    Figure Legend Snippet: Decreased oxidative folding of proinsulin in IRE1α-RNase domain KO MIN6 cells. (A) Levels of Ins genes and transcription factors that regulate Ins genes relative to Gapdh expression levels in the indicated MIN6 cells were measured by qRT-PCR. White bars indicate control MIN6 ( Ire1α fl/fl ) cells (Con). Blue bars indicate MIN6 ( Ire1α fl/fl ) cells infected with Ad-GFP. Red bars indicate MIN6 ( Ire1α ΔR/ΔR ) cells infected with Ad-Cre. (B) To detect newly synthesized preproinsulin and proinsulin in the indicated MIN6 cells, MIN6 cells were pulsed with [ 35 S]Met/Cys for 30 min. Radiolabeled preproinsulin and proinsulin were then immunoprecipitated, separated by NuPAGE, and detected by autoradiography. The ratio of the translation of preproinsulin and proinsulin in MIN6 ( Ire1α ΔR/ΔR ) cells to that of MIN6 ( Ire1α fl/fl ) cells was quantified. Blue bar, MIN6 ( Ire1α fl/fl ) cells; red bar, MIN6 ( Ire1α ΔR/ΔR ) cells. (C) To compare the rate of conversion from preproinsulin to proinsulin in MIN6 ( Ire1α fl/fl or Ire1α ΔR/ΔR ) cells, the cells were pulsed for 1 min with [ 35 S]Met/Cys and then chased for the indicated times. The ratio of proinsulin to the sum of preproinsulin and proinsulin was quantified. The values for MIN6 ( Ire1α fl/fl ) and MIN6 ( Ire1α ΔR/ΔR ) cells are indicated by a blue solid line and by a red broken line, respectively. The values obtained did not differ significantly among time points by Student’s t test. (D) To compare the rate of oxidative folding of proinsulin in MIN6 ( Ire1α fl/fl or Ire1α ΔR/ΔR ) cells, the cells were pulsed for 5 min with [ 35 S]Met/Cys and then chased for the indicated times. Radiolabeled proinsulin was immunoprecipitated with an antiproinsulin antibody, boiled with (Reducing) or without (Nonreducing) DTT, separated by NuPAGE, and detected by autoradiography. The closed diamond indicates preproinsulin. (E) The ratio of oxidized proinsulin (left, red arrowhead) to total proinsulin (right, white arrowhead) in D was quantified. Error bars represent means and SD for each group. n = 3. *, P

    Techniques Used: Expressing, Quantitative RT-PCR, Infection, Synthesized, Immunoprecipitation, Autoradiography

    4) Product Images from "Unexpected Activity of a Novel Kunitz-type Inhibitor"

    Article Title: Unexpected Activity of a Novel Kunitz-type Inhibitor

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M116.724344

    Inhibition of the auto-catalytic activation of FhCL1 by rFhKT1 and rFhKT1Leu 15 /Arg 15 . M, molecular mass marker; lane 1, FhCL1 zymogen; lane 2, mature FhCL1 following auto-catalytic activation at pH 4.5 for 1 h; lane 3, FhCL1 and rFhKT1 following incubation at pH 4.5 for 1 h; lane 4, FhCL1 and rFhKT1Leu 15 /Arg 15 following incubation at pH 4.5 for 1 h; lane 5, FhCL1 and bovine pancreatic trypsin inhibitor following incubation at pH 4.5 for 1 h. Double white arrowhead indicates FhCL1 zymogen; double gray arrowhead indicates mature FhCL1, and double black arrowhead indicates FhCL1 pro-peptide removed following auto-catalytic activation. Single white arrowhead indicates position of FhKT1; single black arrowhead indicates FhKT1Leu 15 /Arg 15 , and single gray arrowhead shows position of BPTI. Proteins were separated on a NuPAGE Novex 4–12% BisTris protein gel and stained with Biosafe Coomassie (Bio-Rad). M, molecular mass markers.
    Figure Legend Snippet: Inhibition of the auto-catalytic activation of FhCL1 by rFhKT1 and rFhKT1Leu 15 /Arg 15 . M, molecular mass marker; lane 1, FhCL1 zymogen; lane 2, mature FhCL1 following auto-catalytic activation at pH 4.5 for 1 h; lane 3, FhCL1 and rFhKT1 following incubation at pH 4.5 for 1 h; lane 4, FhCL1 and rFhKT1Leu 15 /Arg 15 following incubation at pH 4.5 for 1 h; lane 5, FhCL1 and bovine pancreatic trypsin inhibitor following incubation at pH 4.5 for 1 h. Double white arrowhead indicates FhCL1 zymogen; double gray arrowhead indicates mature FhCL1, and double black arrowhead indicates FhCL1 pro-peptide removed following auto-catalytic activation. Single white arrowhead indicates position of FhKT1; single black arrowhead indicates FhKT1Leu 15 /Arg 15 , and single gray arrowhead shows position of BPTI. Proteins were separated on a NuPAGE Novex 4–12% BisTris protein gel and stained with Biosafe Coomassie (Bio-Rad). M, molecular mass markers.

    Techniques Used: Inhibition, Activation Assay, Marker, Incubation, Staining

    Structural representation of FhKT1 and FhKT1Leu 15 /Arg 15 and their recombinant expression. A, sequence alignment of BPTI, FhKT1, and FhKT1Leu 15 /Arg 15 . The asterisk denotes the P1 site at position 15. Lines indicate the conserved disulfide bonds that occur between Cys 1 and Cys 6 , Cys 2 and Cys 4 , and Cys 3 and Cys 5 , with cysteine residues highlighted in black bars. B, recombinant forms of FhKT1 and FhKT1Leu 15 /Arg 15 were expressed as secretory proteins in the methylotrophic yeast P. pastoris with a yield of ∼5–10 mg of soluble protein from each 1 liter of culture. rFhKT1 ( lane 1 ) and rFhKT1Leu 15 /Arg 15 ( lane 2 ) were isolated by NTA-affinity chromatography and analyzed by NuPAGE Novex 4–12% BisTris protein gel, stained with Biosafe Coomassie (Bio-Rad). C, homology model of FhKT1 built based on BPTI (PDB code 3OTJ ) displaying the three disulfide bonds ( yellow ), the α-helix ( red ), and anti-parallel β-sheets ( blue ) characteristic of KT protease inhibitors. The P1 Leu 15 located at the peak of the reactive loop is also shown in gray .
    Figure Legend Snippet: Structural representation of FhKT1 and FhKT1Leu 15 /Arg 15 and their recombinant expression. A, sequence alignment of BPTI, FhKT1, and FhKT1Leu 15 /Arg 15 . The asterisk denotes the P1 site at position 15. Lines indicate the conserved disulfide bonds that occur between Cys 1 and Cys 6 , Cys 2 and Cys 4 , and Cys 3 and Cys 5 , with cysteine residues highlighted in black bars. B, recombinant forms of FhKT1 and FhKT1Leu 15 /Arg 15 were expressed as secretory proteins in the methylotrophic yeast P. pastoris with a yield of ∼5–10 mg of soluble protein from each 1 liter of culture. rFhKT1 ( lane 1 ) and rFhKT1Leu 15 /Arg 15 ( lane 2 ) were isolated by NTA-affinity chromatography and analyzed by NuPAGE Novex 4–12% BisTris protein gel, stained with Biosafe Coomassie (Bio-Rad). C, homology model of FhKT1 built based on BPTI (PDB code 3OTJ ) displaying the three disulfide bonds ( yellow ), the α-helix ( red ), and anti-parallel β-sheets ( blue ) characteristic of KT protease inhibitors. The P1 Leu 15 located at the peak of the reactive loop is also shown in gray .

    Techniques Used: Recombinant, Expressing, Sequencing, Isolation, Affinity Chromatography, Staining

    5) Product Images from "IRE1–XBP1 pathway regulates oxidative proinsulin folding in pancreatic β cells"

    Article Title: IRE1–XBP1 pathway regulates oxidative proinsulin folding in pancreatic β cells

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201707143

    Detection of mixed-disulfide complexes between proinsulin and PDI family members in MIN6 cells and reconstitution of the five PDIs in MIN6 ( Ire1α ΔR/ΔR ) cells. (A) Cys-mediated interaction between PDI family proteins and newly synthesized proinsulin during proinsulin folding. WT MIN6 cells were transiently transfected with indicated FLAG-tagged PDI family proteins, pulsed for 5 min with [ 35 S]Met/Cys, directly treated with TCA, and subjected to alkylation with NEM. Mixed-disulfide complexes between PDIs and proinsulin were collected by consecutive immunoprecipitation (first immunoprecipitation by anti-FLAG antibody; second immunoprecipitation by antiproinsulin antibody). After elution, samples were treated with (Reducing) or without (Nonreducing) DTT, separated by NuPAGE, and detected by autoradiography. (B) MIN6 ( Ire1α ΔR/ΔR ) cells were transiently transfected with WT five PDIs or five CM PDIs. The expression of insulin was induced by culturing cells for 4 h in the presence of low glucose (LG; L; 1.67 mM glucose/KRBH; 4 h) or high glucose (HG; H; 16.7 mM glucose/KRBH; 4 h), and the amounts of secreted insulin were measured by ELISA. White bars indicate insulin secretion upon exposure to low glucose. Black bars indicate insulin secretion upon exposure to high glucose. (C) MIN6 ( Ire1α fl/fl ) cells were transiently transfected with WT PDI/PDIA1 or WT five PDIs. The expression of insulin was induced, and amounts of secreted insulin were measured as described in B. n = 3. **, P
    Figure Legend Snippet: Detection of mixed-disulfide complexes between proinsulin and PDI family members in MIN6 cells and reconstitution of the five PDIs in MIN6 ( Ire1α ΔR/ΔR ) cells. (A) Cys-mediated interaction between PDI family proteins and newly synthesized proinsulin during proinsulin folding. WT MIN6 cells were transiently transfected with indicated FLAG-tagged PDI family proteins, pulsed for 5 min with [ 35 S]Met/Cys, directly treated with TCA, and subjected to alkylation with NEM. Mixed-disulfide complexes between PDIs and proinsulin were collected by consecutive immunoprecipitation (first immunoprecipitation by anti-FLAG antibody; second immunoprecipitation by antiproinsulin antibody). After elution, samples were treated with (Reducing) or without (Nonreducing) DTT, separated by NuPAGE, and detected by autoradiography. (B) MIN6 ( Ire1α ΔR/ΔR ) cells were transiently transfected with WT five PDIs or five CM PDIs. The expression of insulin was induced by culturing cells for 4 h in the presence of low glucose (LG; L; 1.67 mM glucose/KRBH; 4 h) or high glucose (HG; H; 16.7 mM glucose/KRBH; 4 h), and the amounts of secreted insulin were measured by ELISA. White bars indicate insulin secretion upon exposure to low glucose. Black bars indicate insulin secretion upon exposure to high glucose. (C) MIN6 ( Ire1α fl/fl ) cells were transiently transfected with WT PDI/PDIA1 or WT five PDIs. The expression of insulin was induced, and amounts of secreted insulin were measured as described in B. n = 3. **, P

    Techniques Used: Synthesized, Transfection, Immunoprecipitation, Autoradiography, Expressing, Enzyme-linked Immunosorbent Assay

    Decreased oxidative folding of proinsulin in IRE1α-RNase domain KO MIN6 cells. (A) Levels of Ins genes and transcription factors that regulate Ins genes relative to Gapdh expression levels in the indicated MIN6 cells were measured by qRT-PCR. White bars indicate control MIN6 ( Ire1α fl/fl ) cells (Con). Blue bars indicate MIN6 ( Ire1α fl/fl ) cells infected with Ad-GFP. Red bars indicate MIN6 ( Ire1α ΔR/ΔR ) cells infected with Ad-Cre. (B) To detect newly synthesized preproinsulin and proinsulin in the indicated MIN6 cells, MIN6 cells were pulsed with [ 35 S]Met/Cys for 30 min. Radiolabeled preproinsulin and proinsulin were then immunoprecipitated, separated by NuPAGE, and detected by autoradiography. The ratio of the translation of preproinsulin and proinsulin in MIN6 ( Ire1α ΔR/ΔR ) cells to that of MIN6 ( Ire1α fl/fl ) cells was quantified. Blue bar, MIN6 ( Ire1α fl/fl ) cells; red bar, MIN6 ( Ire1α ΔR/ΔR ) cells. (C) To compare the rate of conversion from preproinsulin to proinsulin in MIN6 ( Ire1α fl/fl or Ire1α ΔR/ΔR ) cells, the cells were pulsed for 1 min with [ 35 S]Met/Cys and then chased for the indicated times. The ratio of proinsulin to the sum of preproinsulin and proinsulin was quantified. The values for MIN6 ( Ire1α fl/fl ) and MIN6 ( Ire1α ΔR/ΔR ) cells are indicated by a blue solid line and by a red broken line, respectively. The values obtained did not differ significantly among time points by Student’s t test. (D) To compare the rate of oxidative folding of proinsulin in MIN6 ( Ire1α fl/fl or Ire1α ΔR/ΔR ) cells, the cells were pulsed for 5 min with [ 35 S]Met/Cys and then chased for the indicated times. Radiolabeled proinsulin was immunoprecipitated with an antiproinsulin antibody, boiled with (Reducing) or without (Nonreducing) DTT, separated by NuPAGE, and detected by autoradiography. The closed diamond indicates preproinsulin. (E) The ratio of oxidized proinsulin (left, red arrowhead) to total proinsulin (right, white arrowhead) in D was quantified. Error bars represent means and SD for each group. n = 3. *, P
    Figure Legend Snippet: Decreased oxidative folding of proinsulin in IRE1α-RNase domain KO MIN6 cells. (A) Levels of Ins genes and transcription factors that regulate Ins genes relative to Gapdh expression levels in the indicated MIN6 cells were measured by qRT-PCR. White bars indicate control MIN6 ( Ire1α fl/fl ) cells (Con). Blue bars indicate MIN6 ( Ire1α fl/fl ) cells infected with Ad-GFP. Red bars indicate MIN6 ( Ire1α ΔR/ΔR ) cells infected with Ad-Cre. (B) To detect newly synthesized preproinsulin and proinsulin in the indicated MIN6 cells, MIN6 cells were pulsed with [ 35 S]Met/Cys for 30 min. Radiolabeled preproinsulin and proinsulin were then immunoprecipitated, separated by NuPAGE, and detected by autoradiography. The ratio of the translation of preproinsulin and proinsulin in MIN6 ( Ire1α ΔR/ΔR ) cells to that of MIN6 ( Ire1α fl/fl ) cells was quantified. Blue bar, MIN6 ( Ire1α fl/fl ) cells; red bar, MIN6 ( Ire1α ΔR/ΔR ) cells. (C) To compare the rate of conversion from preproinsulin to proinsulin in MIN6 ( Ire1α fl/fl or Ire1α ΔR/ΔR ) cells, the cells were pulsed for 1 min with [ 35 S]Met/Cys and then chased for the indicated times. The ratio of proinsulin to the sum of preproinsulin and proinsulin was quantified. The values for MIN6 ( Ire1α fl/fl ) and MIN6 ( Ire1α ΔR/ΔR ) cells are indicated by a blue solid line and by a red broken line, respectively. The values obtained did not differ significantly among time points by Student’s t test. (D) To compare the rate of oxidative folding of proinsulin in MIN6 ( Ire1α fl/fl or Ire1α ΔR/ΔR ) cells, the cells were pulsed for 5 min with [ 35 S]Met/Cys and then chased for the indicated times. Radiolabeled proinsulin was immunoprecipitated with an antiproinsulin antibody, boiled with (Reducing) or without (Nonreducing) DTT, separated by NuPAGE, and detected by autoradiography. The closed diamond indicates preproinsulin. (E) The ratio of oxidized proinsulin (left, red arrowhead) to total proinsulin (right, white arrowhead) in D was quantified. Error bars represent means and SD for each group. n = 3. *, P

    Techniques Used: Expressing, Quantitative RT-PCR, Infection, Synthesized, Immunoprecipitation, Autoradiography

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    Transduction:

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    Centrifugation:

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    Blocking Assay:

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    Electrophoresis:

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    Incubation:

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    Article Title:
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    Infection:

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    Mass Spectrometry:

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    BIA-KA:

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    Modification:

    Article Title: Intranasal Administration of Mesenchymal Stem Cells Ameliorates the Abnormal Dopamine Transmission System and Inflammatory Reaction in the R6/2 Mouse Model of Huntington Disease
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    Western Blot:

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    Article Title: Sialylation Controls Prion Fate in Vivo *
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    Derivative Assay:

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    Protease Inhibitor:

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    Cell Culture:

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    Imaging:

    Article Title: Intranasal Administration of Mesenchymal Stem Cells Ameliorates the Abnormal Dopamine Transmission System and Inflammatory Reaction in the R6/2 Mouse Model of Huntington Disease
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    Protein Concentration:

    Article Title: Characterization of the 18 kDa translocator protein (TSPO) expression in post‐mortem normal and Alzheimer's disease brains
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    Sonication:

    Article Title: Intranasal Administration of Mesenchymal Stem Cells Ameliorates the Abnormal Dopamine Transmission System and Inflammatory Reaction in the R6/2 Mouse Model of Huntington Disease
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    Recombinant:

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    Molecular Weight:

    Article Title: Characterization of the 18 kDa translocator protein (TSPO) expression in post‐mortem normal and Alzheimer's disease brains
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    Nucleic Acid Electrophoresis:

    Article Title: Sialylation Controls Prion Fate in Vivo *
    Article Snippet: The resulting pellets were suspended in 1× SDS loading buffer (50 μl for spleen, lymph nodes, brain, and kidneys or 35 μl for liver), heated for 10 min in a boiling water bath, and loaded into NuPAGE Novex 12% Bis-Tris Protein Gels (catalog number NP0343BOX, Thermo Fisher). .. Gel electrophoresis was performed at 170 V for 1 h at room temperature in MES-SDS buffer (catalog number NP0002, Thermo Fisher) in XCell SureLock Mini-Cell (catalog number EI0001, Thermo Fisher).

    Mouse Assay:

    Article Title: Intranasal Administration of Mesenchymal Stem Cells Ameliorates the Abnormal Dopamine Transmission System and Inflammatory Reaction in the R6/2 Mouse Model of Huntington Disease
    Article Snippet: Western Blotting Analysis Mice striatal tissues were homogenized in ice-cold 10 volumes w/v modified RIPA buffer (150 mM sodium chloride, 1.0% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 50 mM Tris, 5 mM EDTA pH 8.0) with Complete Proteinase Inhibitor Cocktail tablets (Sigma-Aldrich, 1873580) with a mechanical homogenizer. .. Protein samples were denatured in Lithium dodecyl sulfate (LDS) buffer (NP0007, Thermo Fisher, Darmstadt, Germany) containing 100 mM DTT and separated using NuPAGE Bis-Tris 12% gel (Thermo Fisher, NP0349BOX).

    Article Title: The protection role of Atg16l1 in CD11c+ dendritic cells in murine colitis
    Article Snippet: BMDCs were derived from Atg16l1f/f and Atg16l1ΔDC mice ( ). .. Cells were plated on 6-well plates at 1500,000 cells/well overnight and were infected with S. typhimurium at a multiplicity of infection (MOI) of 1:20 for 60 min. After stimulation, cells were lysed in LDS sample buffer (Novex, NP0007), boiled, and added onto SDS-polyacrylamide gels (Invitrogen, NP0341BOX).

    Protein Extraction:

    Article Title: Sialylation Controls Prion Fate in Vivo *
    Article Snippet: The tissue homogenates of spleen (5% w/v), mediastinal lymph nodes (2.5% w/v), brain (20% w/v), liver (20% w/v), or kidney (10% w/v) were prepared in M-Per lysis buffer (Mammalian Protein Extraction Reagent, catalog number 78501, Thermo Scientific) in the presence of protease inhibitors (cOmplete, Mini, EDTA-free, catalog number 04693159001, Roche Diagnostics), using a bead beater (Model 2412PS-12W-B30, BioSpec Products,) and 0.1-mm glass beads (catalog number 11079101, BioSpec products). .. The resulting pellets were suspended in 1× SDS loading buffer (50 μl for spleen, lymph nodes, brain, and kidneys or 35 μl for liver), heated for 10 min in a boiling water bath, and loaded into NuPAGE Novex 12% Bis-Tris Protein Gels (catalog number NP0343BOX, Thermo Fisher).

    Article Title: Staurosporine Increases Lentiviral Vector Transduction Efficiency of Human Hematopoietic Stem and Progenitor Cells
    Article Snippet: Cell pellets were lysed using 300 μL mammalian protein extraction reagent (M-PER) (Thermo Scientific #78501) and protease inhibitors (Thermo Fisher #78430) for 10 min with rocking at room temperature. .. Cell debris was removed by centrifugation and the collected supernatant was combined with 4× Laemmli loading buffer (Bio-Rad, #161-0747) containing β-mercaptoethanol and heated at 95°C for 3 min. Twenty microliter samples were loaded on a NuPAGE 12% Bis-Tris gels (Invitrogen, #NP0342BOX) and electrophoresed at 200 V for 45 min.

    Article Title: Melody, an ENU mutation in Caspase 3, alters the catalytic cysteine residue and causes sensorineural hearing loss in mice
    Article Snippet: Paragraph title: Protein extraction and fractionation ... Ten micrograms of each cytoplasmic protein sample were run on a 12% Bis-Tris minigel (Invitrogen, NP0341BOX); the nuclear fraction was discarded.

    Lysis:

    Article Title: Procoagulant and immunogenic properties of melanoma exosomes, microvesicles and apoptotic vesicles
    Article Snippet: Western blotting B16-F1 cells or EV fractions were lysed in lysis buffer (0.02% azide, 150 mM NaCl, 0.25% CHAPS, 0.5% Triton-X100, 100 mM Tris, pH 8.0 plus complete™ protease inhibitor; Roche #11-697-498-001). .. Samples were then run on a NuPAGE 12% Bis-Tris gel (Invitrogen #NP0342BOX) in NuPAGE MOPS-SDS running buffer (NuPAGE #NP0001) plus 0.5 mL of antioxidant (Invitrogen #NP0005) for two hours at 150 V and 126 mA on ice.

    Article Title: Sialylation Controls Prion Fate in Vivo *
    Article Snippet: The tissue homogenates of spleen (5% w/v), mediastinal lymph nodes (2.5% w/v), brain (20% w/v), liver (20% w/v), or kidney (10% w/v) were prepared in M-Per lysis buffer (Mammalian Protein Extraction Reagent, catalog number 78501, Thermo Scientific) in the presence of protease inhibitors (cOmplete, Mini, EDTA-free, catalog number 04693159001, Roche Diagnostics), using a bead beater (Model 2412PS-12W-B30, BioSpec Products,) and 0.1-mm glass beads (catalog number 11079101, BioSpec products). .. The resulting pellets were suspended in 1× SDS loading buffer (50 μl for spleen, lymph nodes, brain, and kidneys or 35 μl for liver), heated for 10 min in a boiling water bath, and loaded into NuPAGE Novex 12% Bis-Tris Protein Gels (catalog number NP0343BOX, Thermo Fisher).

    Article Title: Impaired kidney structure and function in spinal muscular atrophy
    Article Snippet: Western blot Protein was extracted from frozen kidney samples using RIPA lysis buffer (Cell Signaling Technology, Danvers, MA). .. Protein samples (30 μg) were run on a NuPAGE 12% Bis-Tris Protein Gel (NP0342Box, Invitrogen by Thermo Fisher Scientific) electrophoresis and transferred to a nitrocellulose membrane.

    Article Title: Melody, an ENU mutation in Caspase 3, alters the catalytic cysteine residue and causes sensorineural hearing loss in mice
    Article Snippet: E17.5 heads were lysed in 1× lysis buffer containing protease inhibitors using a glass homogenizer. .. Ten micrograms of each cytoplasmic protein sample were run on a 12% Bis-Tris minigel (Invitrogen, NP0341BOX); the nuclear fraction was discarded.

    Purification:

    Article Title: Quantitative immunoassay for mink immunoglobulin in serum and milk
    Article Snippet: .. Specificity analysis by electrophoresis and Western blot The purified mink IgG standard, serum, and milk were mixed with 1/4 LDS sample buffer and 1/10 reducing agent (Life Technologies) before being loaded onto a well on an SDS-PAGE Novex NuPAGE 12% Bis–Tris gel (Ref# NP0341BOX, Invitrogen, Carlsbad, CA, USA). .. NuPAGE MES buffer system was used according to manufacturer’s instructions (Life Technologies) and the gel was run at 200 V constant current.

    Article Title: Quantitative immunoassay for mink immunoglobulin in serum and milk
    Article Snippet: .. The purified mink IgG standard, serum, and milk were mixed with 1/4 LDS sample buffer and 1/10 reducing agent (Life Technologies) before being loaded onto a well on an SDS-PAGE Novex NuPAGE 12% Bis–Tris gel (Ref# NP0341BOX, Invitrogen, Carlsbad, CA, USA). .. NuPAGE MES buffer system was used according to manufacturer’s instructions (Life Technologies) and the gel was run at 200 V constant current.

    SDS Page:

    Article Title: Quantitative immunoassay for mink immunoglobulin in serum and milk
    Article Snippet: .. Specificity analysis by electrophoresis and Western blot The purified mink IgG standard, serum, and milk were mixed with 1/4 LDS sample buffer and 1/10 reducing agent (Life Technologies) before being loaded onto a well on an SDS-PAGE Novex NuPAGE 12% Bis–Tris gel (Ref# NP0341BOX, Invitrogen, Carlsbad, CA, USA). .. NuPAGE MES buffer system was used according to manufacturer’s instructions (Life Technologies) and the gel was run at 200 V constant current.

    Article Title:
    Article Snippet: .. The solvent was exchanged to 50 mM ammonium bicarbonate and further concentrated to 100 µ l. Aliquots of the solution (10 µ l) were mixed with 10 μ l of 4× NuPAGE lithium dodecyl sulfate loading buffer (Invitrogen, Carsbad, CA) by incubation at 95°C for 5 minutes and then subjected to SDS-PAGE on a NuPAGE Novex 10% Bis-Tris mini gel (Invitrogen NP0341BOX). .. After electrophoresis at a constant 180 V for 30 minutes, gels were stained with Coomassie Blue.

    Article Title: Quantitative immunoassay for mink immunoglobulin in serum and milk
    Article Snippet: .. The purified mink IgG standard, serum, and milk were mixed with 1/4 LDS sample buffer and 1/10 reducing agent (Life Technologies) before being loaded onto a well on an SDS-PAGE Novex NuPAGE 12% Bis–Tris gel (Ref# NP0341BOX, Invitrogen, Carlsbad, CA, USA). .. NuPAGE MES buffer system was used according to manufacturer’s instructions (Life Technologies) and the gel was run at 200 V constant current.

    Article Title: Characterization of the 18 kDa translocator protein (TSPO) expression in post‐mortem normal and Alzheimer's disease brains
    Article Snippet: Paragraph title: SDS‐PAGE and Western blot ... 5 μg of protein per temporal cortex or cerebellar sample and 20 μg per frontal cortex sample were loaded into 17‐well 1.0‐mm‐thick NuPAGE 12% Bis‐Tris gels (Thermo Scientific, NP0343BOX).

    Software:

    Article Title: Staurosporine Increases Lentiviral Vector Transduction Efficiency of Human Hematopoietic Stem and Progenitor Cells
    Article Snippet: Cell debris was removed by centrifugation and the collected supernatant was combined with 4× Laemmli loading buffer (Bio-Rad, #161-0747) containing β-mercaptoethanol and heated at 95°C for 3 min. Twenty microliter samples were loaded on a NuPAGE 12% Bis-Tris gels (Invitrogen, #NP0342BOX) and electrophoresed at 200 V for 45 min. .. The bands were visualized using fluorescently tagged secondary antibodies (LI-COR Biosciences, #926-32213 and #926-68070) and imaged on Odyssey CLX using Image Studio 3.1.4 software.

    Binding Assay:

    Article Title: The protection role of Atg16l1 in CD11c+ dendritic cells in murine colitis
    Article Snippet: Cells were plated on 6-well plates at 1500,000 cells/well overnight and were infected with S. typhimurium at a multiplicity of infection (MOI) of 1:20 for 60 min. After stimulation, cells were lysed in LDS sample buffer (Novex, NP0007), boiled, and added onto SDS-polyacrylamide gels (Invitrogen, NP0341BOX). .. Blots were washed and stained with HRP-conjugated secondary antibody (Goat ant-rabbit IgG-HRP, Santa-Cruz, sc-2004), and binding was detected by chemiluminescence (Thermo Scientific, 34080).

    Concentration Assay:

    Article Title: Intranasal Administration of Mesenchymal Stem Cells Ameliorates the Abnormal Dopamine Transmission System and Inflammatory Reaction in the R6/2 Mouse Model of Huntington Disease
    Article Snippet: Protein samples were denatured in Lithium dodecyl sulfate (LDS) buffer (NP0007, Thermo Fisher, Darmstadt, Germany) containing 100 mM DTT and separated using NuPAGE Bis-Tris 12% gel (Thermo Fisher, NP0349BOX). .. Blots were incubated overnight at 4 °C with the following primary antibodies: Anti-pro-brain-derived neurotrophic factor (BDNF) (1:500, Sigma-Aldrich, P1374-200UL), anti-nerve growth factor (NGF) (1:1000 Abcam, ab6199), anti-DARPP-32 (1:5000, Epitomics, 1710-1), anti-tyrosine hydroxylase (TH) at a concentration of 1:1000 (Merck Millipore, AB1542), anti-Iba1 at a concentration of 1:1000 (Wako, 019-1974), and anti-beta actin (1: 5000, Sigma-Aldrich, A5441).

    Article Title:
    Article Snippet: The incubation was started by the addition of a NADPH-regenerating system (final concentration: 1.3 mM NADPH, 4 mM glucose-6-phosophate, 2 U/ml glucose-6-phosphate dehydrogenase, and 3.3 mM potassium phosphate) and allowed to proceed at 37°C for 80 minutes. .. The solvent was exchanged to 50 mM ammonium bicarbonate and further concentrated to 100 µ l. Aliquots of the solution (10 µ l) were mixed with 10 μ l of 4× NuPAGE lithium dodecyl sulfate loading buffer (Invitrogen, Carsbad, CA) by incubation at 95°C for 5 minutes and then subjected to SDS-PAGE on a NuPAGE Novex 10% Bis-Tris mini gel (Invitrogen NP0341BOX).

    Article Title: Sialylation of the prion protein glycans controls prion replication rate and glycoform ratio
    Article Snippet: Western blots 10 μL of 10% brain homogenate or dsPMCAb-derived materials were supplemented with 5 μL 1% SDS and 5 μL proteinase K (cat. # P8107S, New England Biolabs, Ipswich, MA), to a final concentration of 0.25% SDS and 25 μg/mL proteinase K, and incubated at 37 °C for 1 hour. .. Then, SDS sample buffer was added, samples were boiled for 10 minutes and loaded onto NuPage 12% BisTris gel (cat. # NP0341BOX, Life Technologies, Carlsbad, CA), transferred to PVDF membrane, and probed with 3F4 (cat. #SIG-39600, Covance) or Ab3531 antibody (Abcam).

    Fractionation:

    Article Title: Melody, an ENU mutation in Caspase 3, alters the catalytic cysteine residue and causes sensorineural hearing loss in mice
    Article Snippet: Paragraph title: Protein extraction and fractionation ... Ten micrograms of each cytoplasmic protein sample were run on a 12% Bis-Tris minigel (Invitrogen, NP0341BOX); the nuclear fraction was discarded.

    Staining:

    Article Title:
    Article Snippet: The solvent was exchanged to 50 mM ammonium bicarbonate and further concentrated to 100 µ l. Aliquots of the solution (10 µ l) were mixed with 10 μ l of 4× NuPAGE lithium dodecyl sulfate loading buffer (Invitrogen, Carsbad, CA) by incubation at 95°C for 5 minutes and then subjected to SDS-PAGE on a NuPAGE Novex 10% Bis-Tris mini gel (Invitrogen NP0341BOX). .. After electrophoresis at a constant 180 V for 30 minutes, gels were stained with Coomassie Blue.

    Article Title: The protection role of Atg16l1 in CD11c+ dendritic cells in murine colitis
    Article Snippet: Cells were plated on 6-well plates at 1500,000 cells/well overnight and were infected with S. typhimurium at a multiplicity of infection (MOI) of 1:20 for 60 min. After stimulation, cells were lysed in LDS sample buffer (Novex, NP0007), boiled, and added onto SDS-polyacrylamide gels (Invitrogen, NP0341BOX). .. Proteins were transferred to polyvinylidenedifluoride membranes (Fisher-Scientific, 07-200-165), blocked for 60 min with 1% BSA, and stained overnight with the indicated primary antibody (LC3, Cell Signaling, 3868; P62, Sigma-Aldrich, P0067; b-actin, Cell Signaling, 8457; Phospho-p40phox (Thr154) Antibody, Cell Signaling, 4311; Atg16l1 D6D5, Cell Signaling) at 4 °C.

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  • 90
    Thermo Fisher bistris protein gels
    Inhibition of the auto-catalytic activation of FhCL1 by rFhKT1 and rFhKT1Leu 15 /Arg 15 . M, molecular mass marker; lane 1, FhCL1 zymogen; lane 2, mature FhCL1 following auto-catalytic activation at pH 4.5 for 1 h; lane 3, FhCL1 and rFhKT1 following incubation at pH 4.5 for 1 h; lane 4, FhCL1 and rFhKT1Leu 15 /Arg 15 following incubation at pH 4.5 for 1 h; lane 5, FhCL1 and bovine pancreatic trypsin inhibitor following incubation at pH 4.5 for 1 h. Double white arrowhead indicates FhCL1 zymogen; double gray arrowhead indicates mature FhCL1, and double black arrowhead indicates FhCL1 pro-peptide removed following auto-catalytic activation. Single white arrowhead indicates position of FhKT1; single black arrowhead indicates FhKT1Leu 15 /Arg 15 , and single gray arrowhead shows position of BPTI. Proteins were separated on a NuPAGE Novex 4–12% <t>BisTris</t> protein gel and stained with Biosafe Coomassie (Bio-Rad). M, molecular mass markers.
    Bistris Protein Gels, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bistris protein gels/product/Thermo Fisher
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    bistris protein gels - by Bioz Stars, 2020-02
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    90
    Thermo Fisher nupage mops sds buffer kit
    Western blots of purified proteins. (A) Various ST6Gal I and ST3Gal IV purified proteins on Western blots detected by mouse anti-His, HRP-goat anti-mouse IgG with DAB staining. (B) PNGase F digested ST6Gal I (+) and undigested (−) control. Loading was ∼2 μg purified from ProCHO-AT media separated on 12% <t>NuPAGE</t> Novex Bis-Tris gels in <t>MOPS</t> buffer under reducing conditions. Marker: BenchMark His-tagged protein standard. See Table 1 for explanation of all clone names.
    Nupage Mops Sds Buffer Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nupage mops sds buffer kit/product/Thermo Fisher
    Average 90 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    nupage mops sds buffer kit - by Bioz Stars, 2020-02
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    90
    Thermo Fisher nupage bis tris sds page gel
    In vitro translation of dihydrofolate reductase ( DHFR ) in the presence of synthetic peptides representing the toxin and antitoxin proteins of the D11S_1718‐1719 and D11S_1194‐1195 toxin‐antitoxin ( TA ) systems. In vitro translation reactions were incubated with the toxin or antitoxin proteins of the TA systems, either alone or in combination and reaction products were electrophoresed in a <t>Tris‐Glycine</t> <t>SDS</t> ‐ <t>PAGE</t> gel. Lanes 1 and 8, positive control reaction without toxin or antitoxin protein (arrows indicates the DHFR protein); Lane 2, D11S_1194 toxin only; Lane 3, D11S_1195 antitoxin only; Lane 4, equal mixture of the D11S_1194‐1195 toxin and antitoxin; Lane 5, D11S_1718 toxin only; Lane 6, D11S_1719 antitoxin only; Lane 7, equal mixture of the D11S_1718‐1719 toxin and antitoxin; Lane 9, empty; Lane 10, size markers
    Nupage Bis Tris Sds Page Gel, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nupage bis tris sds page gel/product/Thermo Fisher
    Average 90 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    nupage bis tris sds page gel - by Bioz Stars, 2020-02
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    Image Search Results


    Inhibition of the auto-catalytic activation of FhCL1 by rFhKT1 and rFhKT1Leu 15 /Arg 15 . M, molecular mass marker; lane 1, FhCL1 zymogen; lane 2, mature FhCL1 following auto-catalytic activation at pH 4.5 for 1 h; lane 3, FhCL1 and rFhKT1 following incubation at pH 4.5 for 1 h; lane 4, FhCL1 and rFhKT1Leu 15 /Arg 15 following incubation at pH 4.5 for 1 h; lane 5, FhCL1 and bovine pancreatic trypsin inhibitor following incubation at pH 4.5 for 1 h. Double white arrowhead indicates FhCL1 zymogen; double gray arrowhead indicates mature FhCL1, and double black arrowhead indicates FhCL1 pro-peptide removed following auto-catalytic activation. Single white arrowhead indicates position of FhKT1; single black arrowhead indicates FhKT1Leu 15 /Arg 15 , and single gray arrowhead shows position of BPTI. Proteins were separated on a NuPAGE Novex 4–12% BisTris protein gel and stained with Biosafe Coomassie (Bio-Rad). M, molecular mass markers.

    Journal: The Journal of Biological Chemistry

    Article Title: Unexpected Activity of a Novel Kunitz-type Inhibitor

    doi: 10.1074/jbc.M116.724344

    Figure Lengend Snippet: Inhibition of the auto-catalytic activation of FhCL1 by rFhKT1 and rFhKT1Leu 15 /Arg 15 . M, molecular mass marker; lane 1, FhCL1 zymogen; lane 2, mature FhCL1 following auto-catalytic activation at pH 4.5 for 1 h; lane 3, FhCL1 and rFhKT1 following incubation at pH 4.5 for 1 h; lane 4, FhCL1 and rFhKT1Leu 15 /Arg 15 following incubation at pH 4.5 for 1 h; lane 5, FhCL1 and bovine pancreatic trypsin inhibitor following incubation at pH 4.5 for 1 h. Double white arrowhead indicates FhCL1 zymogen; double gray arrowhead indicates mature FhCL1, and double black arrowhead indicates FhCL1 pro-peptide removed following auto-catalytic activation. Single white arrowhead indicates position of FhKT1; single black arrowhead indicates FhKT1Leu 15 /Arg 15 , and single gray arrowhead shows position of BPTI. Proteins were separated on a NuPAGE Novex 4–12% BisTris protein gel and stained with Biosafe Coomassie (Bio-Rad). M, molecular mass markers.

    Article Snippet: A Bradford assay was carried out to determine protein yield, and protein purity was visualized by gel electrophoresis on precast NuPAGE Novex 4–12% BisTris protein gels (ThermoFisher Scientific).

    Techniques: Inhibition, Activation Assay, Marker, Incubation, Staining

    Structural representation of FhKT1 and FhKT1Leu 15 /Arg 15 and their recombinant expression. A, sequence alignment of BPTI, FhKT1, and FhKT1Leu 15 /Arg 15 . The asterisk denotes the P1 site at position 15. Lines indicate the conserved disulfide bonds that occur between Cys 1 and Cys 6 , Cys 2 and Cys 4 , and Cys 3 and Cys 5 , with cysteine residues highlighted in black bars. B, recombinant forms of FhKT1 and FhKT1Leu 15 /Arg 15 were expressed as secretory proteins in the methylotrophic yeast P. pastoris with a yield of ∼5–10 mg of soluble protein from each 1 liter of culture. rFhKT1 ( lane 1 ) and rFhKT1Leu 15 /Arg 15 ( lane 2 ) were isolated by NTA-affinity chromatography and analyzed by NuPAGE Novex 4–12% BisTris protein gel, stained with Biosafe Coomassie (Bio-Rad). C, homology model of FhKT1 built based on BPTI (PDB code 3OTJ ) displaying the three disulfide bonds ( yellow ), the α-helix ( red ), and anti-parallel β-sheets ( blue ) characteristic of KT protease inhibitors. The P1 Leu 15 located at the peak of the reactive loop is also shown in gray .

    Journal: The Journal of Biological Chemistry

    Article Title: Unexpected Activity of a Novel Kunitz-type Inhibitor

    doi: 10.1074/jbc.M116.724344

    Figure Lengend Snippet: Structural representation of FhKT1 and FhKT1Leu 15 /Arg 15 and their recombinant expression. A, sequence alignment of BPTI, FhKT1, and FhKT1Leu 15 /Arg 15 . The asterisk denotes the P1 site at position 15. Lines indicate the conserved disulfide bonds that occur between Cys 1 and Cys 6 , Cys 2 and Cys 4 , and Cys 3 and Cys 5 , with cysteine residues highlighted in black bars. B, recombinant forms of FhKT1 and FhKT1Leu 15 /Arg 15 were expressed as secretory proteins in the methylotrophic yeast P. pastoris with a yield of ∼5–10 mg of soluble protein from each 1 liter of culture. rFhKT1 ( lane 1 ) and rFhKT1Leu 15 /Arg 15 ( lane 2 ) were isolated by NTA-affinity chromatography and analyzed by NuPAGE Novex 4–12% BisTris protein gel, stained with Biosafe Coomassie (Bio-Rad). C, homology model of FhKT1 built based on BPTI (PDB code 3OTJ ) displaying the three disulfide bonds ( yellow ), the α-helix ( red ), and anti-parallel β-sheets ( blue ) characteristic of KT protease inhibitors. The P1 Leu 15 located at the peak of the reactive loop is also shown in gray .

    Article Snippet: A Bradford assay was carried out to determine protein yield, and protein purity was visualized by gel electrophoresis on precast NuPAGE Novex 4–12% BisTris protein gels (ThermoFisher Scientific).

    Techniques: Recombinant, Expressing, Sequencing, Isolation, Affinity Chromatography, Staining

    Western blots of purified proteins. (A) Various ST6Gal I and ST3Gal IV purified proteins on Western blots detected by mouse anti-His, HRP-goat anti-mouse IgG with DAB staining. (B) PNGase F digested ST6Gal I (+) and undigested (−) control. Loading was ∼2 μg purified from ProCHO-AT media separated on 12% NuPAGE Novex Bis-Tris gels in MOPS buffer under reducing conditions. Marker: BenchMark His-tagged protein standard. See Table 1 for explanation of all clone names.

    Journal: PeerJ

    Article Title: Engineering of CHO cells for the production of vertebrate recombinant sialyltransferases

    doi: 10.7717/peerj.5788

    Figure Lengend Snippet: Western blots of purified proteins. (A) Various ST6Gal I and ST3Gal IV purified proteins on Western blots detected by mouse anti-His, HRP-goat anti-mouse IgG with DAB staining. (B) PNGase F digested ST6Gal I (+) and undigested (−) control. Loading was ∼2 μg purified from ProCHO-AT media separated on 12% NuPAGE Novex Bis-Tris gels in MOPS buffer under reducing conditions. Marker: BenchMark His-tagged protein standard. See Table 1 for explanation of all clone names.

    Article Snippet: Proteins were analyzed in reducing conditions on 12% NuPAGE Novex Bis–Tris gels using the NuPAGE MOPS SDS Buffer Kit (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Western Blot, Purification, Staining, Marker

    In vitro translation of dihydrofolate reductase ( DHFR ) in the presence of synthetic peptides representing the toxin and antitoxin proteins of the D11S_1718‐1719 and D11S_1194‐1195 toxin‐antitoxin ( TA ) systems. In vitro translation reactions were incubated with the toxin or antitoxin proteins of the TA systems, either alone or in combination and reaction products were electrophoresed in a Tris‐Glycine SDS ‐ PAGE gel. Lanes 1 and 8, positive control reaction without toxin or antitoxin protein (arrows indicates the DHFR protein); Lane 2, D11S_1194 toxin only; Lane 3, D11S_1195 antitoxin only; Lane 4, equal mixture of the D11S_1194‐1195 toxin and antitoxin; Lane 5, D11S_1718 toxin only; Lane 6, D11S_1719 antitoxin only; Lane 7, equal mixture of the D11S_1718‐1719 toxin and antitoxin; Lane 9, empty; Lane 10, size markers

    Journal: Molecular Oral Microbiology

    Article Title: Identification and functional characterization of type II toxin/antitoxin systems in Aggregatibacter actinomycetemcomitans. Identification and functional characterization of type II toxin/antitoxin systems in Aggregatibacter actinomycetemcomitans

    doi: 10.1111/omi.12215

    Figure Lengend Snippet: In vitro translation of dihydrofolate reductase ( DHFR ) in the presence of synthetic peptides representing the toxin and antitoxin proteins of the D11S_1718‐1719 and D11S_1194‐1195 toxin‐antitoxin ( TA ) systems. In vitro translation reactions were incubated with the toxin or antitoxin proteins of the TA systems, either alone or in combination and reaction products were electrophoresed in a Tris‐Glycine SDS ‐ PAGE gel. Lanes 1 and 8, positive control reaction without toxin or antitoxin protein (arrows indicates the DHFR protein); Lane 2, D11S_1194 toxin only; Lane 3, D11S_1195 antitoxin only; Lane 4, equal mixture of the D11S_1194‐1195 toxin and antitoxin; Lane 5, D11S_1718 toxin only; Lane 6, D11S_1719 antitoxin only; Lane 7, equal mixture of the D11S_1718‐1719 toxin and antitoxin; Lane 9, empty; Lane 10, size markers

    Article Snippet: Synthetic peptides were dissolved in 0.05 m phosphate buffer, pH 7.8, containing 300 mm NaCl and 0.01% trifluoroacetic acid and peptide purity and size was confirmed using a NuPAGE Bis‐Tris SDS‐PAGE gel (Thermo Fisher Scientific, Waltham, MA).

    Techniques: In Vitro, Incubation, SDS Page, Positive Control