polyvinylidene difluoride pvdf membranes  (Roche)


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    Structured Review

    Roche polyvinylidene difluoride pvdf membranes
    In vitro cleavage of WT vSag7 or the CS12X triple mutant by cathepsin L. Equal amounts of purified [ 35 S]methionine-labeled proteins produced with a coupled in vitro transcription-translation system were subjected to cleavage with 10 ng of purified cathepsin L, fractionated by <t>SDS–15%</t> PAGE, transferred to <t>PVDF</t> membranes, and exposed to a PhosphorImager screen. Time of cleavage is in minutes. These data are representative of three independent experiments. The values on the left are the molecular masses in kilodaltons.
    Polyvinylidene Difluoride Pvdf Membranes, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Alternative Proteolytic Processing of Mouse Mammary Tumor Virus Superantigens"

    Article Title: Alternative Proteolytic Processing of Mouse Mammary Tumor Virus Superantigens

    Journal: Journal of Virology

    doi:

    In vitro cleavage of WT vSag7 or the CS12X triple mutant by cathepsin L. Equal amounts of purified [ 35 S]methionine-labeled proteins produced with a coupled in vitro transcription-translation system were subjected to cleavage with 10 ng of purified cathepsin L, fractionated by SDS–15% PAGE, transferred to PVDF membranes, and exposed to a PhosphorImager screen. Time of cleavage is in minutes. These data are representative of three independent experiments. The values on the left are the molecular masses in kilodaltons.
    Figure Legend Snippet: In vitro cleavage of WT vSag7 or the CS12X triple mutant by cathepsin L. Equal amounts of purified [ 35 S]methionine-labeled proteins produced with a coupled in vitro transcription-translation system were subjected to cleavage with 10 ng of purified cathepsin L, fractionated by SDS–15% PAGE, transferred to PVDF membranes, and exposed to a PhosphorImager screen. Time of cleavage is in minutes. These data are representative of three independent experiments. The values on the left are the molecular masses in kilodaltons.

    Techniques Used: In Vitro, Mutagenesis, Purification, Labeling, Produced, Polyacrylamide Gel Electrophoresis

    In vitro cleavage of vSag7 mutants by recombinant convertases. vSags were produced with a coupled in vitro transcription-and-translation system as [ 35 S]methionine-labeled proteins. Equal amounts of Ni-nitrilotriacetic acid-purified material were subjected to cleavage with equal units of convertase activity. The mutated and WT vSags were digested overnight. Cleavage products were separated by SDS–15% PAGE, transferred to PVDF membranes, and exposed on PhosphorImager screens. (A) Expected molecular weights of cleavage products. (B) Cleavage of the CS2X mutant. (C) Cleavage of the CS1X mutant. (D) Cleavage of WT vSag7. (E) Cleavage of the CS12 mutant. Lanes: 1, furin; 2, PC5; 3, PC7; 4, furin plus EDTA; 5, uncleaved control. These data are representative of three independent experiments. The values shown are the molecular masses in kilodaltons.
    Figure Legend Snippet: In vitro cleavage of vSag7 mutants by recombinant convertases. vSags were produced with a coupled in vitro transcription-and-translation system as [ 35 S]methionine-labeled proteins. Equal amounts of Ni-nitrilotriacetic acid-purified material were subjected to cleavage with equal units of convertase activity. The mutated and WT vSags were digested overnight. Cleavage products were separated by SDS–15% PAGE, transferred to PVDF membranes, and exposed on PhosphorImager screens. (A) Expected molecular weights of cleavage products. (B) Cleavage of the CS2X mutant. (C) Cleavage of the CS1X mutant. (D) Cleavage of WT vSag7. (E) Cleavage of the CS12 mutant. Lanes: 1, furin; 2, PC5; 3, PC7; 4, furin plus EDTA; 5, uncleaved control. These data are representative of three independent experiments. The values shown are the molecular masses in kilodaltons.

    Techniques Used: In Vitro, Recombinant, Produced, Labeling, Purification, Activity Assay, Polyacrylamide Gel Electrophoresis, Mutagenesis

    Related Articles

    Nucleic Acid Electrophoresis:

    Article Title: Deficient O-GlcNAc Glycosylation Impairs Regulatory T Cell Differentiation and Notch Signaling in Autoimmune Hepatitis
    Article Snippet: .. After centrifugation, protein samples were subjected to 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto polyvinylidene fluoride (PVDF) membranes (Roche, Mannheim, Germany). .. The membranes were then blocked in TBST (1 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.05% Tween-20) containing 5% skim milk for 60 min, and incubated overnight at 4°C with diluted primary antibodies against GAPDH, β-actin, NALP2 and EOGT (Abcam, Database link: ), as well as cleaved caspase 3, cleaved caspase 7 and RBPJ (Cell Signaling Technology).

    Centrifugation:

    Article Title: Deficient O-GlcNAc Glycosylation Impairs Regulatory T Cell Differentiation and Notch Signaling in Autoimmune Hepatitis
    Article Snippet: .. After centrifugation, protein samples were subjected to 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto polyvinylidene fluoride (PVDF) membranes (Roche, Mannheim, Germany). .. The membranes were then blocked in TBST (1 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.05% Tween-20) containing 5% skim milk for 60 min, and incubated overnight at 4°C with diluted primary antibodies against GAPDH, β-actin, NALP2 and EOGT (Abcam, Database link: ), as well as cleaved caspase 3, cleaved caspase 7 and RBPJ (Cell Signaling Technology).

    Blocking Assay:

    Article Title: Changes in the Muscarinic Receptors on the Colonic Smooth Muscles of Rats with Spinal Cord Injury
    Article Snippet: .. The proteins were transferred to 0.45 µm polyvinylidene difluoride (PVDF) membranes (Roche Diagnostics GmbH, Mannheim, Germany) using protein transfer apparatus (mini Transblot cell; Bio-Rad Laboratories) at 100 V for 90 min. To prevent non-specific binding of the primary antibody, the PVDF membranes were incubated with blocking buffer (5% skim milk in 0.05% tween 20-Tris buffer saline [TBST, pH 7.6]) for 1 h. The membranes were washed 3 times with TBST and incubated overnight at 4℃ with primary antibodies (PGP 9.5, HSP 27, and RhoA) that were diluted to 1:1,000 with TBS containing 3% BSA. ..

    Incubation:

    Article Title: Changes in the Muscarinic Receptors on the Colonic Smooth Muscles of Rats with Spinal Cord Injury
    Article Snippet: .. The proteins were transferred to 0.45 µm polyvinylidene difluoride (PVDF) membranes (Roche Diagnostics GmbH, Mannheim, Germany) using protein transfer apparatus (mini Transblot cell; Bio-Rad Laboratories) at 100 V for 90 min. To prevent non-specific binding of the primary antibody, the PVDF membranes were incubated with blocking buffer (5% skim milk in 0.05% tween 20-Tris buffer saline [TBST, pH 7.6]) for 1 h. The membranes were washed 3 times with TBST and incubated overnight at 4℃ with primary antibodies (PGP 9.5, HSP 27, and RhoA) that were diluted to 1:1,000 with TBS containing 3% BSA. ..

    Article Title: SETMAR functions in illegitimate DNA recombination and non-homologous end joining
    Article Snippet: .. Proteins were transferred to a polyvinylidene difluoride (PVDF) membrane, which was blocked in 5% milk or BSA (Roche) and incubated with specific primary antibodies at 4°C overnight. .. After washing, membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies for one hour at room temperature, washed, and signals were detected with the ECL system (Promega) and Fuji medical X-ray film (Fujifilm).

    Polyacrylamide Gel Electrophoresis:

    Article Title: Alternative Proteolytic Processing of Mouse Mammary Tumor Virus Superantigens
    Article Snippet: .. Products were fractionated by sodium dodecyl sulfate (SDS)–15% polyacrylamide gel electrophoresis (PAGE), transferred to polyvinylidene difluoride (PVDF) membranes (Roche Diagnostics), and exposed overnight on PhosphorImager screens (Molecular Dynamics, Sunnyvale, Calif.). ..

    Affinity Purification:

    Article Title: The Recruitment of the Saccharomyces cerevisiae Paf1 Complex to Active Genes Requires a Domain of Rtf1 That Directly Interacts with the Spt4-Spt5 Complex
    Article Snippet: .. Proteins were resolved on SDS-polyacrylamide gels, transferred to nitrocellulose or polyvinylidene difluoride (PVDF) membranes, and probed with primary antibodies against Rtf1 , the hemagglutinin (HA) epitope (1:2,500 dilution; Roche), total histone H3 (1:30,000 dilution) , trimethylated H3 K4 (H3 K4 Me3) (39159, 1:2,000 dilution; Active Motif), H3 K4 Me2 (07-030, 1:2,000 dilution; Upstate), H3 K79 Me2/3 (ab2621, 1:2,000 dilution; Abcam), Paf1 (1:1,000 dilution; gift from Judith Jaehning), glucose-6-phosphate dehydrogenase (G6PDH) (A9521, 1:30,000 dilution; Sigma), tobacco etch virus (TEV) protease-cleaved tandem affinity purification (TAP) tag (CAB 1001, 1:2,500; Thermo Scientific), or an antibody that detects the uncleaved TAP tag (P1291, peroxidase-antiperoxidase, 1:2,000 dilution; Sigma). .. After incubation with the primary antibodies, membranes were probed with sheep anti-mouse or donkey anti-rabbit secondary antibodies (1:5,000 dilution; GE Healthcare) and visualized using enhanced chemiluminescence substrate (PerkinElmer).

    Binding Assay:

    Article Title: Changes in the Muscarinic Receptors on the Colonic Smooth Muscles of Rats with Spinal Cord Injury
    Article Snippet: .. The proteins were transferred to 0.45 µm polyvinylidene difluoride (PVDF) membranes (Roche Diagnostics GmbH, Mannheim, Germany) using protein transfer apparatus (mini Transblot cell; Bio-Rad Laboratories) at 100 V for 90 min. To prevent non-specific binding of the primary antibody, the PVDF membranes were incubated with blocking buffer (5% skim milk in 0.05% tween 20-Tris buffer saline [TBST, pH 7.6]) for 1 h. The membranes were washed 3 times with TBST and incubated overnight at 4℃ with primary antibodies (PGP 9.5, HSP 27, and RhoA) that were diluted to 1:1,000 with TBS containing 3% BSA. ..

    SDS Page:

    Article Title: Salmonella produce microRNA-like RNA fragment Sal-1 in the infected cells to facilitate intracellular survival
    Article Snippet: .. The proteins (40 μg) were separated by SDS-PAGE and electrotransferred to polyvinylidene difluoride (PVDF) membranes (Roche, Indianapolis, IN). ..

    Article Title: TRIM52 inhibits Japanese Encephalitis Virus replication by degrading the viral NS2A
    Article Snippet: .. Equal amounts of proteins were separated using 12% sodium dodecyl sulfate polyacrylamide gels (SDS-PAGE) and transferred onto polyvinylidene fluoride (PVDF) membranes (Roche, UK). ..

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    Roche pvdf western blotting membranes
    ClfB binds to Fg A α -chain. (a) Ligand affinity blotting. Human Fg was fractionated by <t>SDS-PAGE</t> and stained with Coomassie blue (lane 1), or transferred to a <t>PVDF-membrane</t> and probed with rClfB 45–542 (lane 2) or rClfA 220–559 (lane 3). Binding of rClf proteins was detected with anti-Clf polyclonal antisera. (b) Adherence of S. aureus Newman ClfA − to immobilized recombinant A α - (•), B β - (▴) or γ - (▾) chains of Fg. (c) Adherence of S. aureus Newman ClfA − (○) and Newman ClfA − ClfB − (•) to immobilized recombinant A α -chain of Fg. Cells were grown to exponential phase and suspensions (1×10 8 c.f.u.) were added to wells. Adherent bacteria were detected by crystal violet staining. The experiment was performed twice with similar results and values represent means± sd of triplicate wells.
    Pvdf Western Blotting Membranes, supplied by Roche, used in various techniques. Bioz Stars score: 88/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Roche pvdf membranes
    The C-terminal domain of the isolated PT β-subunit lacks an ER retention signal. COS-7 cells were transfected with cDNAs coding for EGFP control, wild-type, and mutant HA-PT β-subunit with alanine substitution of the potential ER retention motif (HA-PT β RRR→AAA) or deletion of the entire C-terminal tail (HA-PT β ΔCT). Total cell extracts were prepared, incubated in the absence (−) or presence (+) of endo H or <t>PNGase</t> F and analyzed by HA Western blotting ( WB ). After electrotransfer of the proteins, <t>PVDF</t> membranes were probed with antibodies against HA followed by incubation with HRP-coupled secondary antibodies and ECL analysis. Positions of the N -glycosylated PT β-subunit and of the molecular mass marker proteins are indicated.
    Pvdf Membranes, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 428 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pvdf membranes/product/Roche
    Average 94 stars, based on 428 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    ClfB binds to Fg A α -chain. (a) Ligand affinity blotting. Human Fg was fractionated by SDS-PAGE and stained with Coomassie blue (lane 1), or transferred to a PVDF-membrane and probed with rClfB 45–542 (lane 2) or rClfA 220–559 (lane 3). Binding of rClf proteins was detected with anti-Clf polyclonal antisera. (b) Adherence of S. aureus Newman ClfA − to immobilized recombinant A α - (•), B β - (▴) or γ - (▾) chains of Fg. (c) Adherence of S. aureus Newman ClfA − (○) and Newman ClfA − ClfB − (•) to immobilized recombinant A α -chain of Fg. Cells were grown to exponential phase and suspensions (1×10 8 c.f.u.) were added to wells. Adherent bacteria were detected by crystal violet staining. The experiment was performed twice with similar results and values represent means± sd of triplicate wells.

    Journal: Microbiology

    Article Title: Identification of the Staphylococcus aureus MSCRAMM clumping factor B (ClfB) binding site in the αC-domain of human fibrinogen

    doi: 10.1099/mic.0.2007/010868-0

    Figure Lengend Snippet: ClfB binds to Fg A α -chain. (a) Ligand affinity blotting. Human Fg was fractionated by SDS-PAGE and stained with Coomassie blue (lane 1), or transferred to a PVDF-membrane and probed with rClfB 45–542 (lane 2) or rClfA 220–559 (lane 3). Binding of rClf proteins was detected with anti-Clf polyclonal antisera. (b) Adherence of S. aureus Newman ClfA − to immobilized recombinant A α - (•), B β - (▴) or γ - (▾) chains of Fg. (c) Adherence of S. aureus Newman ClfA − (○) and Newman ClfA − ClfB − (•) to immobilized recombinant A α -chain of Fg. Cells were grown to exponential phase and suspensions (1×10 8 c.f.u.) were added to wells. Adherent bacteria were detected by crystal violet staining. The experiment was performed twice with similar results and values represent means± sd of triplicate wells.

    Article Snippet: Human Fg (Calbiochem) was separated by SDS-PAGE and transferred electrophoretically to PVDF Western blotting membranes (Roche Applied Science) by the wet system (Bio-Rad) in Tris/HCl (0.02 M), glycine (0.15 M) and methanol (20 %, v/v).

    Techniques: SDS Page, Staining, Binding Assay, Recombinant

    The C-terminal domain of the isolated PT β-subunit lacks an ER retention signal. COS-7 cells were transfected with cDNAs coding for EGFP control, wild-type, and mutant HA-PT β-subunit with alanine substitution of the potential ER retention motif (HA-PT β RRR→AAA) or deletion of the entire C-terminal tail (HA-PT β ΔCT). Total cell extracts were prepared, incubated in the absence (−) or presence (+) of endo H or PNGase F and analyzed by HA Western blotting ( WB ). After electrotransfer of the proteins, PVDF membranes were probed with antibodies against HA followed by incubation with HRP-coupled secondary antibodies and ECL analysis. Positions of the N -glycosylated PT β-subunit and of the molecular mass marker proteins are indicated.

    Journal: The Journal of Biological Chemistry

    Article Title: Transport of the GlcNAc-1-phosphotransferase ?/?-Subunit Precursor Protein to the Golgi Apparatus Requires a Combinatorial Sorting Motif *

    doi: 10.1074/jbc.M112.407676

    Figure Lengend Snippet: The C-terminal domain of the isolated PT β-subunit lacks an ER retention signal. COS-7 cells were transfected with cDNAs coding for EGFP control, wild-type, and mutant HA-PT β-subunit with alanine substitution of the potential ER retention motif (HA-PT β RRR→AAA) or deletion of the entire C-terminal tail (HA-PT β ΔCT). Total cell extracts were prepared, incubated in the absence (−) or presence (+) of endo H or PNGase F and analyzed by HA Western blotting ( WB ). After electrotransfer of the proteins, PVDF membranes were probed with antibodies against HA followed by incubation with HRP-coupled secondary antibodies and ECL analysis. Positions of the N -glycosylated PT β-subunit and of the molecular mass marker proteins are indicated.

    Article Snippet: Peptide N -glycosidase F (PNGase F) and PVDF membranes were from Roche Applied Science, and endoglycosidase H (endo H) was from New England Biolabs (Frankfurt, Germany).

    Techniques: Isolation, Transfection, Mutagenesis, Incubation, Western Blot, Electrotransfer, Marker

    Conditional intestinal epithelial HDAC1/2 loss disrupts epithelial barrier function. A . Total protein extracts from three to five control (Ctrl) or conditional intestinal epithelial HDAC1/2 (HDAC1/2ΔIEC) colons were separated on a 10% SDS-PAGE gel, transferred to a PVDF membrane and analysed by Western blot for expression of Claudin 3 (MW: 23.3 kD) and actin as a loading control. The histograms indicate the ratio of band intensities normalized to actin. Quantification of band intensity was performed with the Quantity One software. Results represent the mean ± SEM (*p≤0.05). B . To measure intestinal permeability, blood was recovered 3 h after gavage of 4-kDa FITC-labeled dextran (n=6). FITC serum concentrations were determined with a RF-5301PC spectrofluorometer (Shimadzu Scientific Instruments, Columbia, MD, USA). Results represent the mean ± SEM (*p≤0.05). C . Total protein extracts from four to five control (Ctrl) or conditional intestinal epithelial HDAC1/2 (HDAC1/2ΔIEC) colons were separated on a 10% SDS-PAGE gel, transferred to a PVDF membrane and analysed by Western blot for expression of Phospho-Stat3 and total Stat3 (MW: 88 kD). The histogram indicates the ratio of Phospho-Stat3 band intensities normalized to Stat3. Quantification of band intensity was performed with the Quantity One software. Results represent the mean ± SEM (*p≤0.05).

    Journal: PLoS ONE

    Article Title: HDAC1 and HDAC2 Restrain the Intestinal Inflammatory Response by Regulating Intestinal Epithelial Cell Differentiation

    doi: 10.1371/journal.pone.0073785

    Figure Lengend Snippet: Conditional intestinal epithelial HDAC1/2 loss disrupts epithelial barrier function. A . Total protein extracts from three to five control (Ctrl) or conditional intestinal epithelial HDAC1/2 (HDAC1/2ΔIEC) colons were separated on a 10% SDS-PAGE gel, transferred to a PVDF membrane and analysed by Western blot for expression of Claudin 3 (MW: 23.3 kD) and actin as a loading control. The histograms indicate the ratio of band intensities normalized to actin. Quantification of band intensity was performed with the Quantity One software. Results represent the mean ± SEM (*p≤0.05). B . To measure intestinal permeability, blood was recovered 3 h after gavage of 4-kDa FITC-labeled dextran (n=6). FITC serum concentrations were determined with a RF-5301PC spectrofluorometer (Shimadzu Scientific Instruments, Columbia, MD, USA). Results represent the mean ± SEM (*p≤0.05). C . Total protein extracts from four to five control (Ctrl) or conditional intestinal epithelial HDAC1/2 (HDAC1/2ΔIEC) colons were separated on a 10% SDS-PAGE gel, transferred to a PVDF membrane and analysed by Western blot for expression of Phospho-Stat3 and total Stat3 (MW: 88 kD). The histogram indicates the ratio of Phospho-Stat3 band intensities normalized to Stat3. Quantification of band intensity was performed with the Quantity One software. Results represent the mean ± SEM (*p≤0.05).

    Article Snippet: 30 µg of total protein extracts were loaded on a 10% or a 15% SDS-polyacrylamide gel and electroblotted on a PVDF membrane (Roche Molecular Biochemicals, Laval, QC, Canada).

    Techniques: SDS Page, Western Blot, Expressing, Software, Permeability, Labeling

    Conditional intestinal epithelial HDAC1/2 loss leads to altered activation of cell homeostasis regulators. Total protein extracts from three to four one-year-old control (Ctrl) or conditional intestinal epithelial HDAC1/2 (HDAC1/2ΔIEC) colons were separated on a 10% SDS-PAGE gel, transferred to a PVDF membrane and analysed by Western blot for expression of ( A ) Cyclin D (Ccnd1, MW: 33.4 kD; Ccnd2, MW: 32.9 kD), cleaved Notch1 (MW: 110 kD) and actin (MW: 41.7 kD) as a loading control; ( B ) phosphorylated and total ribosomal protein S6 (MW: 28.7 kD); ( C ) cleaved caspase 3 (MW: 17 kD), with actin as a loading control. The histograms indicate the ratio of band intensities normalized to actin ( A , C ) or total ribosomal protein S6 ( B ). Quantification of band intensity was performed with the Quantity One software. Results represent the mean ± SEM (*p≤0.05; **p≤0.01). D . Cyclin D1 (Ccnd1) increased expression was confirmed by qPCR analysis of total RNAs isolated from control or conditional intestinal epithelial HDAC1/2 colons. Results represent the mean ± SEM (*p≤0.05).

    Journal: PLoS ONE

    Article Title: HDAC1 and HDAC2 Restrain the Intestinal Inflammatory Response by Regulating Intestinal Epithelial Cell Differentiation

    doi: 10.1371/journal.pone.0073785

    Figure Lengend Snippet: Conditional intestinal epithelial HDAC1/2 loss leads to altered activation of cell homeostasis regulators. Total protein extracts from three to four one-year-old control (Ctrl) or conditional intestinal epithelial HDAC1/2 (HDAC1/2ΔIEC) colons were separated on a 10% SDS-PAGE gel, transferred to a PVDF membrane and analysed by Western blot for expression of ( A ) Cyclin D (Ccnd1, MW: 33.4 kD; Ccnd2, MW: 32.9 kD), cleaved Notch1 (MW: 110 kD) and actin (MW: 41.7 kD) as a loading control; ( B ) phosphorylated and total ribosomal protein S6 (MW: 28.7 kD); ( C ) cleaved caspase 3 (MW: 17 kD), with actin as a loading control. The histograms indicate the ratio of band intensities normalized to actin ( A , C ) or total ribosomal protein S6 ( B ). Quantification of band intensity was performed with the Quantity One software. Results represent the mean ± SEM (*p≤0.05; **p≤0.01). D . Cyclin D1 (Ccnd1) increased expression was confirmed by qPCR analysis of total RNAs isolated from control or conditional intestinal epithelial HDAC1/2 colons. Results represent the mean ± SEM (*p≤0.05).

    Article Snippet: 30 µg of total protein extracts were loaded on a 10% or a 15% SDS-polyacrylamide gel and electroblotted on a PVDF membrane (Roche Molecular Biochemicals, Laval, QC, Canada).

    Techniques: Activation Assay, SDS Page, Western Blot, Expressing, Software, Real-time Polymerase Chain Reaction, Isolation

    C3H/10T1/2 cells were differentiated in the presence of DMSO (D) or 15 µM CX-4945 (CX) and harvested at given time points during differentiation. Proteins were extracted and separated on a 12.5% SDS-polyacrylamide gel and transferred to a PVDF-membrane. C/EBPβ, C/EBPδ, C/EBPα, and PPARγ2 were detected with specific antibodies. GAPDH was used as a loading control.

    Journal: Pharmaceuticals

    Article Title: Inhibition of Protein Kinase CK2 Prevents Adipogenic Differentiation of Mesenchymal Stem Cells Like C3H/10T1/2 Cells

    doi: 10.3390/ph10010022

    Figure Lengend Snippet: C3H/10T1/2 cells were differentiated in the presence of DMSO (D) or 15 µM CX-4945 (CX) and harvested at given time points during differentiation. Proteins were extracted and separated on a 12.5% SDS-polyacrylamide gel and transferred to a PVDF-membrane. C/EBPβ, C/EBPδ, C/EBPα, and PPARγ2 were detected with specific antibodies. GAPDH was used as a loading control.

    Article Snippet: Proteins dissolved in sample buffer (130 mM Tris-HCl, pH 6.8, 0.02% bromophenol blue, 10% β-mercaptoethanol, 20% glycerol, 4% SDS) were separated on a 12.5% SDS- polyacrylamide gel in electrophoresis buffer (25 mM Tris-HCl, pH 8.8, 192 mM glycine, 0.1% SDS) and transferred onto a PVDF membrane (Roche Diagnostics, Mannheim, Germany) in a buffer containing 20 mM Tris-HCl, 150 mM glycine, pH 8.3.

    Techniques: