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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 6598 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
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    97/100 stars

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    Knock-Out:

    Article Title: Partial characterization of the proteome of the mouse striatum.
    Article Snippet: Many diseases of the mammalian CNS, including Parkinson’s (PD) and Lesch Nyhan disease (LND), are associated with programmatic neurodegeneration or dysfunction of dopaminergic neurons in the mesencephalon, the nigrostriatal pathway, and its projections in the striatum [1– 4].. Proteomic studies on brain tissue of both animal models and human PD patients have provided evidence for dysfunction and damage of many pathways, including oxidative stress-related damage, ubiquitin-proteasome dysfunction, mitochondrial energy metabolism deficiencies, and synaptic function [5–11].. To date no such proteomic studies have been reported in the related and rare basal ganglia disorder LND, a developmental rather than a neurodegenerative neurological disorder caused by deficiency of the enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) that regulates a major step in the purine salvage pathway [12].

    Electrofocusing:

    Article Title: Partial characterization of the proteome of the mouse striatum.
    Article Snippet: Many diseases of the mammalian CNS, including Parkinson’s (PD) and Lesch Nyhan disease (LND), are associated with programmatic neurodegeneration or dysfunction of dopaminergic neurons in the mesencephalon, the nigrostriatal pathway, and its projections in the striatum [1– 4].. Proteomic studies on brain tissue of both animal models and human PD patients have provided evidence for dysfunction and damage of many pathways, including oxidative stress-related damage, ubiquitin-proteasome dysfunction, mitochondrial energy metabolism deficiencies, and synaptic function [5–11].. To date no such proteomic studies have been reported in the related and rare basal ganglia disorder LND, a developmental rather than a neurodegenerative neurological disorder caused by deficiency of the enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) that regulates a major step in the purine salvage pathway [12].

    SDS Page:

    Article Title: Partial characterization of the proteome of the mouse striatum.
    Article Snippet: Many diseases of the mammalian CNS, including Parkinson’s (PD) and Lesch Nyhan disease (LND), are associated with programmatic neurodegeneration or dysfunction of dopaminergic neurons in the mesencephalon, the nigrostriatal pathway, and its projections in the striatum [1– 4].. Proteomic studies on brain tissue of both animal models and human PD patients have provided evidence for dysfunction and damage of many pathways, including oxidative stress-related damage, ubiquitin-proteasome dysfunction, mitochondrial energy metabolism deficiencies, and synaptic function [5–11].. To date no such proteomic studies have been reported in the related and rare basal ganglia disorder LND, a developmental rather than a neurodegenerative neurological disorder caused by deficiency of the enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) that regulates a major step in the purine salvage pathway [12].



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    Bio-Rad de lysis solution
    De Lysis Solution, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad 2-de rehydration buffer
    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).
    2 De Rehydration Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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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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    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