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de lysis solution  (Bio-Rad)


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    Structured Review

    Bio-Rad de lysis solution
    De Lysis Solution, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 97/100, based on 6594 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/2+de+buffer/Lysis+Buffer/pm39174378-75-13-27
    Average 97 stars, based on 6594 article reviews
    de lysis solution - by Bioz Stars, 2026-09
    97/100 stars

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    Related Articles

    Lysis:

    Article Title: Short autoinhibitory sequences control phase separation of an essential bacterial transcription termination factor.
    Article Snippet: .. The supernatant was loaded onto 1 mL NiNTA agarose (Qiagen) previously washed with Lysis buffer and applied on a Polyprep column (Bio-Rad) for gravity flow chromatography. ..

    Article Title: Rap1b Activates Endosomal AC9 to Drive the Second cAMP Wave
    Article Snippet: Purified GST–RalGDS–RBD bound to glutathione–Sepharose beads (10 μg) was added and incubated with the supernatants for 60 min at 4 °C with rotation. .. Beads were washed four times with lysis buffer, resuspended in Laemmli sample buffer (1610737, Bio–Rad), and analyzed by SDS–PAGE (12%) followed by transfer to PVDF membranes (EMD Millipore, IPVH00010) for immunoblotting. .. His–tagged proteins (His–GFP, His–C1a, and His–C2a; 50 μg) were expressed in E. coli and captured on 40 μL of 50% Ni–NTA agarose beads (Qiagen, 30210) by rotation at 4 °C for 1 h. Beads were briefly centrifuged and washed three times with lysis buffer (25 mM Tris–HCl, pH 7.5, 150 mM NaCl, 5 mM MgCl2, 5% glycerol, 1% Nonidet P–40).

    Article Title: Calciprotein particles disrupt autophagy in vascular endothelial cells and smooth muscle cells
    Article Snippet: .. HAoECs and HAoSMCs were collected in Laemmli sample buffer (Bio-Rad), supplemented with β-mercaptoethanol (5%), to facilitate cell lysis. ..

    Article Title: A single PA-X mutation in bovine-origin H5N1 influenza virus reduces pathogenicity in mice
    Article Snippet: .. Briefly, cell lysates prepared in NP-40 lysis buffer were mixed with SDS-PAGE loading buffer, boiled, resolved by SDS-PAGE (12%), and transferred to nitrocellulose membranes (Bio-Rad). ..

    Article Title: α/Sulfonyl-γ-AApeptide foldamers mitigate Alzheimer's disease pathology by stabilizing transient helical domains in Aβ.
    Article Snippet: Immunoblotting was performed using the Abby system (ProteinSimple), an automated capillary-based Western blot platform that enables size-based protein separation and chemiluminescent detection. .. For in vitro samples (primary neurons treated with Aβ with or without M4 treatment) and in vivo tissue lysates, proteins were extracted using standard lysis buffer, clarified by centrifugation, and quantified using a protein assay kit (Bio-Rad). ..

    Chromatography:

    Article Title: Short autoinhibitory sequences control phase separation of an essential bacterial transcription termination factor.
    Article Snippet: .. The supernatant was loaded onto 1 mL NiNTA agarose (Qiagen) previously washed with Lysis buffer and applied on a Polyprep column (Bio-Rad) for gravity flow chromatography. ..

    Western Blot:

    Article Title: Rap1b Activates Endosomal AC9 to Drive the Second cAMP Wave
    Article Snippet: Purified GST–RalGDS–RBD bound to glutathione–Sepharose beads (10 μg) was added and incubated with the supernatants for 60 min at 4 °C with rotation. .. Beads were washed four times with lysis buffer, resuspended in Laemmli sample buffer (1610737, Bio–Rad), and analyzed by SDS–PAGE (12%) followed by transfer to PVDF membranes (EMD Millipore, IPVH00010) for immunoblotting. .. His–tagged proteins (His–GFP, His–C1a, and His–C2a; 50 μg) were expressed in E. coli and captured on 40 μL of 50% Ni–NTA agarose beads (Qiagen, 30210) by rotation at 4 °C for 1 h. Beads were briefly centrifuged and washed three times with lysis buffer (25 mM Tris–HCl, pH 7.5, 150 mM NaCl, 5 mM MgCl2, 5% glycerol, 1% Nonidet P–40).

    Article Title: Development of novel monoclonal antibodies for detection of pan-Lassa virus.
    Article Snippet: .. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and WB Cell lysate containing NP was boiled at 95°C for 5 minutes with Laemmli buffer (Bio-Rad) and separated in 4–20% Mini-PROTEAN TGX Precast Protein Gels (Bio-Rad). .. At 48 hours post-transfection, cells were washed with PBS and lysed overnight in 4°C using cell lysis buffer consist of 50mM Tris pH 8.0, 300mM NaCl, 0.5% Triton X-100, supplemented with protease inhibitor cocktail (Thermo scientific).

    Staining:

    Article Title: Dependencies in heterogeneous, lineage plastic patient–derived prostate cancer organoids revealed through integrated single–cell multiomics and CRISPR screening
    Article Snippet: Protein concentration was determined using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific). .. Equal amounts of protein were mixed with 1× Laemmli sample buffer (Bio-Rad) and boiled for 10 min before separation by SDS–PAGE on 4–15% Mini-PROTEAN TGX Stain-Free gels (Bio-Rad). .. Proteins were transferred onto PVDF membranes (Bio-Rad), and a Full Range Rainbow recombinant protein ladder (Cytiva) was used as a molecular weight marker.

    Nucleic Acid Electrophoresis:

    Article Title: Development of novel monoclonal antibodies for detection of pan-Lassa virus.
    Article Snippet: .. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and WB Cell lysate containing NP was boiled at 95°C for 5 minutes with Laemmli buffer (Bio-Rad) and separated in 4–20% Mini-PROTEAN TGX Precast Protein Gels (Bio-Rad). .. At 48 hours post-transfection, cells were washed with PBS and lysed overnight in 4°C using cell lysis buffer consist of 50mM Tris pH 8.0, 300mM NaCl, 0.5% Triton X-100, supplemented with protease inhibitor cocktail (Thermo scientific).

    Blocking Assay:

    Article Title: FGFR2-responsive self-assembled hydrogel releases its inhibitor to suppress the progression of endometriosis.
    Article Snippet: Fibroblast growth factor receptor 2 (FGFR2) has emerged as a highly promising therapeutic target due to its critical role in the pathogenesis and progression of endometriosis.. To date, a variety of FGFR2 inhibitors have exhibited remarkable therapeutic efficacy against endometriosis in both preclinical studies and clinical trials.. However, these small-molecule inhibitors suffer from drawbacks including poor targeting ability and high toxicity, and their direct administration has consequently limited their further clinical applications.

    Incubation:

    Article Title: FGFR2-responsive self-assembled hydrogel releases its inhibitor to suppress the progression of endometriosis.
    Article Snippet: Fibroblast growth factor receptor 2 (FGFR2) has emerged as a highly promising therapeutic target due to its critical role in the pathogenesis and progression of endometriosis.. To date, a variety of FGFR2 inhibitors have exhibited remarkable therapeutic efficacy against endometriosis in both preclinical studies and clinical trials.. However, these small-molecule inhibitors suffer from drawbacks including poor targeting ability and high toxicity, and their direct administration has consequently limited their further clinical applications.

    SDS Page:

    Article Title: A single PA-X mutation in bovine-origin H5N1 influenza virus reduces pathogenicity in mice
    Article Snippet: .. Briefly, cell lysates prepared in NP-40 lysis buffer were mixed with SDS-PAGE loading buffer, boiled, resolved by SDS-PAGE (12%), and transferred to nitrocellulose membranes (Bio-Rad). ..

    In Vitro:

    Article Title: α/Sulfonyl-γ-AApeptide foldamers mitigate Alzheimer's disease pathology by stabilizing transient helical domains in Aβ.
    Article Snippet: Immunoblotting was performed using the Abby system (ProteinSimple), an automated capillary-based Western blot platform that enables size-based protein separation and chemiluminescent detection. .. For in vitro samples (primary neurons treated with Aβ with or without M4 treatment) and in vivo tissue lysates, proteins were extracted using standard lysis buffer, clarified by centrifugation, and quantified using a protein assay kit (Bio-Rad). ..

    In Vivo:

    Article Title: α/Sulfonyl-γ-AApeptide foldamers mitigate Alzheimer's disease pathology by stabilizing transient helical domains in Aβ.
    Article Snippet: Immunoblotting was performed using the Abby system (ProteinSimple), an automated capillary-based Western blot platform that enables size-based protein separation and chemiluminescent detection. .. For in vitro samples (primary neurons treated with Aβ with or without M4 treatment) and in vivo tissue lysates, proteins were extracted using standard lysis buffer, clarified by centrifugation, and quantified using a protein assay kit (Bio-Rad). ..

    Centrifugation:

    Article Title: α/Sulfonyl-γ-AApeptide foldamers mitigate Alzheimer's disease pathology by stabilizing transient helical domains in Aβ.
    Article Snippet: Immunoblotting was performed using the Abby system (ProteinSimple), an automated capillary-based Western blot platform that enables size-based protein separation and chemiluminescent detection. .. For in vitro samples (primary neurons treated with Aβ with or without M4 treatment) and in vivo tissue lysates, proteins were extracted using standard lysis buffer, clarified by centrifugation, and quantified using a protein assay kit (Bio-Rad). ..



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    Effect of glucose on the mRNA and protein expression of TvLEGU-2. ( A ) qPCR with specific primers for the tvlegu-2 and α-tubulin genes using 100 ng of cDNA from parasites grown in glucose restriction (GR) and high-glucose (HG) conditions. α-tubulin was used as a normalizing gene; ns, no significant differences. ( B ) Western blot assay of PREs from parasites grown under GR (<1 mM) and HG (50 mM). Coomassie Brilliant-Blue (CBB)-stained 10% SDS PAGE gel for PREs from parasites grown under GR and HG conditions (Lanes 2 and 3), respectively. For WB assays, duplicate gels were transferred onto NC membranes and incubated with different antibodies: Rα-TvLEGU-2pep (1:500 dilution) to detect the TvLEGU-2 protein, Rα-TvCP2r (1:6000 dilution) to detect a control protein overexpressed under GR conditions, Rα-TvTIM (1:1000 dilution) to detect an overexpressed TvTIM control protein under HG conditions, and a negative control with PI serum or no primary antibody (−). Arrowheads show the position of the native TvLEGU-2 (∼55, ∼49, ∼34, and ∼29 kDa) protein bands. ( C ) Silver-stained <t>2-DE</t> protease-rich extracts from parasites grown in normal glucose conditions (25 mM) ( Ca ). WB of duplicate gels transferred onto NC membranes incubated with Rα-TvLEGU-1r (1:1000 dilution) antibody ( Cc ) to serve as a control for the specificity of Rα-TvLEGU-2pep antibody against other legumain proteins, Rα-TvLEGU-2pep (1:100 dilution) ( Cd ) to detect the TvLEGU-2 protein in PREs, or a negative control with PI serum or no primary antibody (−) ( Cb ). Arrowheads show the position of the native TvLEGU-2 (∼34 and ∼29 kDa) proteins. ( D ) Coomassie Brilliant Blue-stained, purified recombinant TvLEGU-1r protein (CBB; Lane 1). WB assays of TvLEGU-1r incubated with Rα-TvLEGU-1r (1:3000 dilution) (Lane 3), or Rα-TvLEGU-2pep (1:1000 dilution) (Lane 4) antibody or PI serum or only the secondary antibody as a negative control (−) (Lane 2). Arrowhead points to the recombinant protein band TvLEGU-1r (∼46 kDa). ( E ) Coomassie Brilliant Blue-stained, purified recombinant TvLEGU-2r protein (CBB; Lane 1). WB assays of TvLEGU-2r incubated with Rα-TvLEGU-2pep (1:1000 dilution) (Lane 4), or Rα-TvLEGU-1r (1:3000 dilution) (Lane 3) antibody, or PI serum or only the secondary antibody as a negative control (−) (Lane 2). Arrowhead points to the recombinant protein band TvLEGU-2r (∼85 kDa). kDa, molecular weight markers in kilodaltons (Bio-Rad).
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    The <t>2-DE</t> proteome images of control (A) and RC-treated (B) platelets. The differentially expressed protein spots were shown by the arrows.
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    Figure 1 Identification of the BaMV-induced proteins in the S100 fraction. A, Genome of BaMV. B, Schematic representations of infectious con- structs which were BaMV-derived in this study. pKn, a binary vector which can express foreign genes driven by the upstream double 35S promoter of cauliflower mosaic virus. It also has a nopaline synthase (Nos) terminator. pKB, which has an infectious full-length BaMV construct in the pKn vector. pKB GFP, has a GFP ORF downstream of TGBp3, which was driven by the same promoter as CP. pKB GFP-2a-CP, GFP, and CP are linked with a 2a protein. <t>C,</t> <t>2-DE</t> analysis of S100 fractions from vector- or BaMV-inoculated N. benthamiana. S100 fractions were extracted at 7 dpi. The proteins from S100 were separated by 2-DE analysis and the excised differentiated dots (indicated as numbers) were analyzed by LC–MS/MS. Agrobacterium tumefaciens bearing pKn or pKB plasmid was infiltrated into 28-d-old N. benthamiana. The infiltrated leaves were collected at 7 dpi. D, Total RNA was extracted from infiltrated leaves and mRNA levels of NbVDACs were quantified by RT-qPCR. Data are mean ± standard de- viation (SD) from at least three independent experiments with three replicates each (N 5 3, n = 3) Asterisks indicate statistically significant differ- ences between the indicated groups by Student’s t test (**P 5 0.01, ***P 5 0.001).
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    Figure 1 Identification of the BaMV-induced proteins in the S100 fraction. A, Genome of BaMV. B, Schematic representations of infectious con- structs which were BaMV-derived in this study. pKn, a binary vector which can express foreign genes driven by the upstream double 35S promoter of cauliflower mosaic virus. It also has a nopaline synthase (Nos) terminator. pKB, which has an infectious full-length BaMV construct in the pKn vector. pKB GFP, has a GFP ORF downstream of TGBp3, which was driven by the same promoter as CP. pKB GFP-2a-CP, GFP, and CP are linked with a 2a protein. <t>C,</t> <t>2-DE</t> analysis of S100 fractions from vector- or BaMV-inoculated N. benthamiana. S100 fractions were extracted at 7 dpi. The proteins from S100 were separated by 2-DE analysis and the excised differentiated dots (indicated as numbers) were analyzed by LC–MS/MS. Agrobacterium tumefaciens bearing pKn or pKB plasmid was infiltrated into 28-d-old N. benthamiana. The infiltrated leaves were collected at 7 dpi. D, Total RNA was extracted from infiltrated leaves and mRNA levels of NbVDACs were quantified by RT-qPCR. Data are mean ± standard de- viation (SD) from at least three independent experiments with three replicates each (N 5 3, n = 3) Asterisks indicate statistically significant differ- ences between the indicated groups by Student’s t test (**P 5 0.01, ***P 5 0.001).
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    Figure 1 Identification of the BaMV-induced proteins in the S100 fraction. A, Genome of BaMV. B, Schematic representations of infectious con- structs which were BaMV-derived in this study. pKn, a binary vector which can express foreign genes driven by the upstream double 35S promoter of cauliflower mosaic virus. It also has a nopaline synthase (Nos) terminator. pKB, which has an infectious full-length BaMV construct in the pKn vector. pKB GFP, has a GFP ORF downstream of TGBp3, which was driven by the same promoter as CP. pKB GFP-2a-CP, GFP, and CP are linked with a 2a protein. <t>C,</t> <t>2-DE</t> analysis of S100 fractions from vector- or BaMV-inoculated N. benthamiana. S100 fractions were extracted at 7 dpi. The proteins from S100 were separated by 2-DE analysis and the excised differentiated dots (indicated as numbers) were analyzed by LC–MS/MS. Agrobacterium tumefaciens bearing pKn or pKB plasmid was infiltrated into 28-d-old N. benthamiana. The infiltrated leaves were collected at 7 dpi. D, Total RNA was extracted from infiltrated leaves and mRNA levels of NbVDACs were quantified by RT-qPCR. Data are mean ± standard de- viation (SD) from at least three independent experiments with three replicates each (N 5 3, n = 3) Asterisks indicate statistically significant differ- ences between the indicated groups by Student’s t test (**P 5 0.01, ***P 5 0.001).
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    Figure 1 Identification of the BaMV-induced proteins in the S100 fraction. A, Genome of BaMV. B, Schematic representations of infectious con- structs which were BaMV-derived in this study. pKn, a binary vector which can express foreign genes driven by the upstream double 35S promoter of cauliflower mosaic virus. It also has a nopaline synthase (Nos) terminator. pKB, which has an infectious full-length BaMV construct in the pKn vector. pKB GFP, has a GFP ORF downstream of TGBp3, which was driven by the same promoter as CP. pKB GFP-2a-CP, GFP, and CP are linked with a 2a protein. <t>C,</t> <t>2-DE</t> analysis of S100 fractions from vector- or BaMV-inoculated N. benthamiana. S100 fractions were extracted at 7 dpi. The proteins from S100 were separated by 2-DE analysis and the excised differentiated dots (indicated as numbers) were analyzed by LC–MS/MS. Agrobacterium tumefaciens bearing pKn or pKB plasmid was infiltrated into 28-d-old N. benthamiana. The infiltrated leaves were collected at 7 dpi. D, Total RNA was extracted from infiltrated leaves and mRNA levels of NbVDACs were quantified by RT-qPCR. Data are mean ± standard de- viation (SD) from at least three independent experiments with three replicates each (N 5 3, n = 3) Asterisks indicate statistically significant differ- ences between the indicated groups by Student’s t test (**P 5 0.01, ***P 5 0.001).
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    Figure 1 Identification of the BaMV-induced proteins in the S100 fraction. A, Genome of BaMV. B, Schematic representations of infectious con- structs which were BaMV-derived in this study. pKn, a binary vector which can express foreign genes driven by the upstream double 35S promoter of cauliflower mosaic virus. It also has a nopaline synthase (Nos) terminator. pKB, which has an infectious full-length BaMV construct in the pKn vector. pKB GFP, has a GFP ORF downstream of TGBp3, which was driven by the same promoter as CP. pKB GFP-2a-CP, GFP, and CP are linked with a 2a protein. <t>C,</t> <t>2-DE</t> analysis of S100 fractions from vector- or BaMV-inoculated N. benthamiana. S100 fractions were extracted at 7 dpi. The proteins from S100 were separated by 2-DE analysis and the excised differentiated dots (indicated as numbers) were analyzed by LC–MS/MS. Agrobacterium tumefaciens bearing pKn or pKB plasmid was infiltrated into 28-d-old N. benthamiana. The infiltrated leaves were collected at 7 dpi. D, Total RNA was extracted from infiltrated leaves and mRNA levels of NbVDACs were quantified by RT-qPCR. Data are mean ± standard de- viation (SD) from at least three independent experiments with three replicates each (N 5 3, n = 3) Asterisks indicate statistically significant differ- ences between the indicated groups by Student’s t test (**P 5 0.01, ***P 5 0.001).
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    Image Search Results


    Effect of glucose on the mRNA and protein expression of TvLEGU-2. ( A ) qPCR with specific primers for the tvlegu-2 and α-tubulin genes using 100 ng of cDNA from parasites grown in glucose restriction (GR) and high-glucose (HG) conditions. α-tubulin was used as a normalizing gene; ns, no significant differences. ( B ) Western blot assay of PREs from parasites grown under GR (<1 mM) and HG (50 mM). Coomassie Brilliant-Blue (CBB)-stained 10% SDS PAGE gel for PREs from parasites grown under GR and HG conditions (Lanes 2 and 3), respectively. For WB assays, duplicate gels were transferred onto NC membranes and incubated with different antibodies: Rα-TvLEGU-2pep (1:500 dilution) to detect the TvLEGU-2 protein, Rα-TvCP2r (1:6000 dilution) to detect a control protein overexpressed under GR conditions, Rα-TvTIM (1:1000 dilution) to detect an overexpressed TvTIM control protein under HG conditions, and a negative control with PI serum or no primary antibody (−). Arrowheads show the position of the native TvLEGU-2 (∼55, ∼49, ∼34, and ∼29 kDa) protein bands. ( C ) Silver-stained 2-DE protease-rich extracts from parasites grown in normal glucose conditions (25 mM) ( Ca ). WB of duplicate gels transferred onto NC membranes incubated with Rα-TvLEGU-1r (1:1000 dilution) antibody ( Cc ) to serve as a control for the specificity of Rα-TvLEGU-2pep antibody against other legumain proteins, Rα-TvLEGU-2pep (1:100 dilution) ( Cd ) to detect the TvLEGU-2 protein in PREs, or a negative control with PI serum or no primary antibody (−) ( Cb ). Arrowheads show the position of the native TvLEGU-2 (∼34 and ∼29 kDa) proteins. ( D ) Coomassie Brilliant Blue-stained, purified recombinant TvLEGU-1r protein (CBB; Lane 1). WB assays of TvLEGU-1r incubated with Rα-TvLEGU-1r (1:3000 dilution) (Lane 3), or Rα-TvLEGU-2pep (1:1000 dilution) (Lane 4) antibody or PI serum or only the secondary antibody as a negative control (−) (Lane 2). Arrowhead points to the recombinant protein band TvLEGU-1r (∼46 kDa). ( E ) Coomassie Brilliant Blue-stained, purified recombinant TvLEGU-2r protein (CBB; Lane 1). WB assays of TvLEGU-2r incubated with Rα-TvLEGU-2pep (1:1000 dilution) (Lane 4), or Rα-TvLEGU-1r (1:3000 dilution) (Lane 3) antibody, or PI serum or only the secondary antibody as a negative control (−) (Lane 2). Arrowhead points to the recombinant protein band TvLEGU-2r (∼85 kDa). kDa, molecular weight markers in kilodaltons (Bio-Rad).

    Journal: Pathogens

    Article Title: Trichomonas vaginalis Legumain-2, TvLEGU-2, Is an Immunogenic Cysteine Peptidase Expressed during Trichomonal Infection

    doi: 10.3390/pathogens13020119

    Figure Lengend Snippet: Effect of glucose on the mRNA and protein expression of TvLEGU-2. ( A ) qPCR with specific primers for the tvlegu-2 and α-tubulin genes using 100 ng of cDNA from parasites grown in glucose restriction (GR) and high-glucose (HG) conditions. α-tubulin was used as a normalizing gene; ns, no significant differences. ( B ) Western blot assay of PREs from parasites grown under GR (<1 mM) and HG (50 mM). Coomassie Brilliant-Blue (CBB)-stained 10% SDS PAGE gel for PREs from parasites grown under GR and HG conditions (Lanes 2 and 3), respectively. For WB assays, duplicate gels were transferred onto NC membranes and incubated with different antibodies: Rα-TvLEGU-2pep (1:500 dilution) to detect the TvLEGU-2 protein, Rα-TvCP2r (1:6000 dilution) to detect a control protein overexpressed under GR conditions, Rα-TvTIM (1:1000 dilution) to detect an overexpressed TvTIM control protein under HG conditions, and a negative control with PI serum or no primary antibody (−). Arrowheads show the position of the native TvLEGU-2 (∼55, ∼49, ∼34, and ∼29 kDa) protein bands. ( C ) Silver-stained 2-DE protease-rich extracts from parasites grown in normal glucose conditions (25 mM) ( Ca ). WB of duplicate gels transferred onto NC membranes incubated with Rα-TvLEGU-1r (1:1000 dilution) antibody ( Cc ) to serve as a control for the specificity of Rα-TvLEGU-2pep antibody against other legumain proteins, Rα-TvLEGU-2pep (1:100 dilution) ( Cd ) to detect the TvLEGU-2 protein in PREs, or a negative control with PI serum or no primary antibody (−) ( Cb ). Arrowheads show the position of the native TvLEGU-2 (∼34 and ∼29 kDa) proteins. ( D ) Coomassie Brilliant Blue-stained, purified recombinant TvLEGU-1r protein (CBB; Lane 1). WB assays of TvLEGU-1r incubated with Rα-TvLEGU-1r (1:3000 dilution) (Lane 3), or Rα-TvLEGU-2pep (1:1000 dilution) (Lane 4) antibody or PI serum or only the secondary antibody as a negative control (−) (Lane 2). Arrowhead points to the recombinant protein band TvLEGU-1r (∼46 kDa). ( E ) Coomassie Brilliant Blue-stained, purified recombinant TvLEGU-2r protein (CBB; Lane 1). WB assays of TvLEGU-2r incubated with Rα-TvLEGU-2pep (1:1000 dilution) (Lane 4), or Rα-TvLEGU-1r (1:3000 dilution) (Lane 3) antibody, or PI serum or only the secondary antibody as a negative control (−) (Lane 2). Arrowhead points to the recombinant protein band TvLEGU-2r (∼85 kDa). kDa, molecular weight markers in kilodaltons (Bio-Rad).

    Article Snippet: For 2-DE assays, PRE was obtained from parasites (6 × 10 7 ) lysed directly in 2-DE rehydration buffer (Bio-Rad, Hercules, CA, USA) in the absence of proteinase inhibitors.

    Techniques: Expressing, Western Blot, Staining, SDS Page, Incubation, Negative Control, Purification, Recombinant, Molecular Weight

    The 2-DE proteome images of control (A) and RC-treated (B) platelets. The differentially expressed protein spots were shown by the arrows.

    Journal: Pharmaceutical Biology

    Article Title: Differential proteomic analysis of platelets suggested target-related proteins in rabbit platelets treated with Rhizoma Corydalis

    doi: 10.1080/13880209.2016.1229340

    Figure Lengend Snippet: The 2-DE proteome images of control (A) and RC-treated (B) platelets. The differentially expressed protein spots were shown by the arrows.

    Article Snippet: Two-dimensional electrophoresis analysis was performed on a GE 2-DE system.

    Techniques:

    Figure 1 Identification of the BaMV-induced proteins in the S100 fraction. A, Genome of BaMV. B, Schematic representations of infectious con- structs which were BaMV-derived in this study. pKn, a binary vector which can express foreign genes driven by the upstream double 35S promoter of cauliflower mosaic virus. It also has a nopaline synthase (Nos) terminator. pKB, which has an infectious full-length BaMV construct in the pKn vector. pKB GFP, has a GFP ORF downstream of TGBp3, which was driven by the same promoter as CP. pKB GFP-2a-CP, GFP, and CP are linked with a 2a protein. C, 2-DE analysis of S100 fractions from vector- or BaMV-inoculated N. benthamiana. S100 fractions were extracted at 7 dpi. The proteins from S100 were separated by 2-DE analysis and the excised differentiated dots (indicated as numbers) were analyzed by LC–MS/MS. Agrobacterium tumefaciens bearing pKn or pKB plasmid was infiltrated into 28-d-old N. benthamiana. The infiltrated leaves were collected at 7 dpi. D, Total RNA was extracted from infiltrated leaves and mRNA levels of NbVDACs were quantified by RT-qPCR. Data are mean ± standard de- viation (SD) from at least three independent experiments with three replicates each (N 5 3, n = 3) Asterisks indicate statistically significant differ- ences between the indicated groups by Student’s t test (**P 5 0.01, ***P 5 0.001).

    Journal: Plant physiology

    Article Title: Voltage-dependent anion channel proteins associate with dynamic Bamboo mosaic virus-induced complexes.

    doi: 10.1093/plphys/kiab519

    Figure Lengend Snippet: Figure 1 Identification of the BaMV-induced proteins in the S100 fraction. A, Genome of BaMV. B, Schematic representations of infectious con- structs which were BaMV-derived in this study. pKn, a binary vector which can express foreign genes driven by the upstream double 35S promoter of cauliflower mosaic virus. It also has a nopaline synthase (Nos) terminator. pKB, which has an infectious full-length BaMV construct in the pKn vector. pKB GFP, has a GFP ORF downstream of TGBp3, which was driven by the same promoter as CP. pKB GFP-2a-CP, GFP, and CP are linked with a 2a protein. C, 2-DE analysis of S100 fractions from vector- or BaMV-inoculated N. benthamiana. S100 fractions were extracted at 7 dpi. The proteins from S100 were separated by 2-DE analysis and the excised differentiated dots (indicated as numbers) were analyzed by LC–MS/MS. Agrobacterium tumefaciens bearing pKn or pKB plasmid was infiltrated into 28-d-old N. benthamiana. The infiltrated leaves were collected at 7 dpi. D, Total RNA was extracted from infiltrated leaves and mRNA levels of NbVDACs were quantified by RT-qPCR. Data are mean ± standard de- viation (SD) from at least three independent experiments with three replicates each (N 5 3, n = 3) Asterisks indicate statistically significant differ- ences between the indicated groups by Student’s t test (**P 5 0.01, ***P 5 0.001).

    Article Snippet: The pellet was washed 3 times with methanol and resolved in 2-DE rehydration buffer (7 M urea, 2 M thiourea, 4% CHAPS, 1% IPG-buffer and 60 mM DTT) and separated by 13 cm Immobiline DryStrip pH 3–10 (GE healthcare, Chicago, IL, USA) with the conditions 500 V for 1 h, 1,000 V for 1 h, 2,000 V for 1 h, 8,000 V for 2.5 h with gradient and 8,000 V for 2 h. The strip was soaked in equilibration buffer I (50 mM Tris–HCl pH 8.8, 6 M urea, 30% glycerol, 2% sodium dodecyl sulfate (SDS) and 1% dithiothreitol) for 15 min and transferred to equilibration buffer II (50 mM Tris–HCl pH 8.8, 6M urea, 30% glycerol, 2% SDS, and 135 mM iodoacetamide) for the other 15 min. Next, the strips were placed onto 10–16% gradient SDS–polyacrylamide gels sealed by a 1% low-melting point agarose solution containing 1% agarose, 25 mM Tris base, 192 mM glycine, 0.1% SDS, and 0.002% (wt/vol) bromophenol blue.

    Techniques: Derivative Assay, Plasmid Preparation, Virus, Construct, Liquid Chromatography with Mass Spectroscopy, Quantitative RT-PCR