monoclonal anti beta actin (Millipore)

Name:
Monoclonal Anti beta Actin antibody
Description:
Actin is a highly conserved protein that is a major component of both the cytoskeletal and contractile structures in all cell types It varies in amount being related to the type of differentiation and to the functional state of cells and tissues Actin can be found in two different forms of aggregation the globular or the fibrillar state and at least six distinct isoforms occur in vertebrates The actins exhibit over 90 sequence homology but each isoform has a unique NH2 terminal sequence The isoforms are comprised of three alpha actins skeletal cardiac smooth one beta actin beta non muscle and two gamma actins gamma smooth muscle and gamma non muscle Recognizes an epitope located on the N terminal end of the beta isoform of actin It specifically labels beta actin in a wide variety of tissues and species In immunofluorescent staining of chicken gizzard ultrathin cryosections the antibody labels the dense bodies the longitudinal channels linking consecutive dense bodies that are also occupied by desmin and the membrane associated dense plaque The antibody does not react with adult cardiac or skeletal muscle besides traces due to contamination of the sample with non muscle cells or with beta actin expressing cells in Dictyostelium discoideum amoeba The epitope recognized by the antibody is resistant to formalin fixation and paraffin embedding B5 methacarn ethanol or Bouin s solutions may be also used as fixatives The antibody can be used as probes for beta actin as an internal control in immunoblotting
Catalog Number:
A5316
Molecular Weight:
antigen mol wt 42 kDa
Price:
None
Applications:
Mouse Monoclonal Anti-β-Actin antibody has been used for western blot analyses. The antibody can also be used for immunohistochemistry, indirect immunofluorescence (1:1,000) using cultured human or chicken fibroblasts, and indirect ELISA.Western blot analysis of MDCK cell lysates were performed using monoclonal anti-actin antibody as a primary antibody.Monoclonal mouse anti-actin antibody was used as a loading control for western blot analysis of immunoprecipitated proteins from rat dorsal root ganglion cocultures.
Host:
mouse
Conjugate:
unconjugated
Immunogen:
slightly modified β-cytoplasmic actin N-terminal peptide, Ac-Asp-Asp-Asp-Ile-Ala-Ala-Leu-Val-Ile-Asp-Asn-Gly-Ser-Gly-Lys, conjugated to KLH.
Category:
Antibodies
Isotype:
IgG2a
Source:
goat
Reactivity:
human
Score:
85
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Structured Review

Actin is a highly conserved protein that is a major component of both the cytoskeletal and contractile structures in all cell types It varies in amount being related to the type of differentiation and to the functional state of cells and tissues Actin can be found in two different forms of aggregation the globular or the fibrillar state and at least six distinct isoforms occur in vertebrates The actins exhibit over 90 sequence homology but each isoform has a unique NH2 terminal sequence The isoforms are comprised of three alpha actins skeletal cardiac smooth one beta actin beta non muscle and two gamma actins gamma smooth muscle and gamma non muscle Recognizes an epitope located on the N terminal end of the beta isoform of actin It specifically labels beta actin in a wide variety of tissues and species In immunofluorescent staining of chicken gizzard ultrathin cryosections the antibody labels the dense bodies the longitudinal channels linking consecutive dense bodies that are also occupied by desmin and the membrane associated dense plaque The antibody does not react with adult cardiac or skeletal muscle besides traces due to contamination of the sample with non muscle cells or with beta actin expressing cells in Dictyostelium discoideum amoeba The epitope recognized by the antibody is resistant to formalin fixation and paraffin embedding B5 methacarn ethanol or Bouin s solutions may be also used as fixatives The antibody can be used as probes for beta actin as an internal control in immunoblotting
https://www.bioz.com/result/monoclonal anti beta actin/product/Millipore
Average 99 stars, based on 2 article reviews
Price from $9.99 to $1999.99
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Clone Assay:Article Title: LC3B is not recruited along with the autophagy elongation complex (ATG5-12/16L1) at HCV replication site and is dispensable for viral replication Article Snippet: hATG5 and hATG16L1 sequences were cloned into peGFP-C1 plasmid (Clontech) to form pGFP-ATG5 and pGFP-ATG16L1, respectively. .. Rabbit polyclonal anti-LC3, rabbit polyclonal anti-ATG5 (used for western blot), mouse monoclonal anti-Flag, and Centrifugation:Article Title: Generation and characterisation of a parkin-Pacrg knockout mouse line and a Pacrg knockout mouse line Article Snippet: PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 (A5441, Sigma-Aldrich). .. PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 ( Article Title: Attenuation of autophagy flux by 6-shogaol sensitizes human liver cancer cells to TRAIL-induced apoptosis via p53 and ROS Article Snippet: The β-actin antibody was from Sigma-Aldrich (cat. no. A2228; 1:2,000, Merck KGaA, Darmstadt, Germany), antibodies against microtubule-associated proteins 1A/1B light chain 3B (LC3)-I/II (cat. no. 3868; 1:1,000), cleaved caspase-3 (cas3; cat. no. 9661; 1:500) and p62 (cat. no. 5114; 1:1,000) were from Cell Signaling Technology, Inc., the p53 (cat. no. sc-6243; 1:1,000) antibody was from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and the caspase-8 (cas8; cat. no. 551242; 1:1,000) antibody was from BD Biosciences (Franklin Lakes, NJ, USA). .. The Article Title: Post-ischemic estradiol treatment reduced glial response and triggers distinct cortical and hippocampal signaling in a rat model of cerebral ischemia Article Snippet: Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), β-actin (1:1,000; clone AC-15, Sigma no. A5441), GSK3 (1:1,000; Biosource no. 44–610), phospho-GSK3Ser21/9 (1:1,000; Cell Signaling no. 9331), SAPK-JNK (1:1,000; Cell Signaling no. 9252), pSAPK-JNKThr183/Tyr185 (1:1,000; Cell Signaling no. 9251). .. Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), Article Title: Luteolin Induces microRNA-132 Expression and Modulates Neurite Outgrowth in PC12 Cells Article Snippet: The cellular debris was removed by centrifugation (8,000×g for 15 min) at 4°C, and the cell lysate was carefully transferred to a microcentrifuge tube. .. The membranes were blocked at 4°C in PBST blocking buffer (1% non-fat dried milk in PBS containing 0.1% Tween-20) for 8 h. Blots were incubated with the appropriate antibodies overnight at 4°C: anti-CREB (1∶1000), anti-phospho-CREB (Ser-133) (1∶1000), anti-ERK (1∶1000 ), anti-phospho-ERK (1∶1000 ) (Cell Signaling Technology, Inc.), and Autoradiography:Article Title: Post-ischemic estradiol treatment reduced glial response and triggers distinct cortical and hippocampal signaling in a rat model of cerebral ischemia Article Snippet: Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), Blocking Assay:Article Title: Generation and characterisation of a parkin-Pacrg knockout mouse line and a Pacrg knockout mouse line Article Snippet: PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 (A5441, Sigma-Aldrich). .. PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 ( Article Title: Deleterious Effect of Advanced CKD on Glyoxalase System Activity not Limited to Diabetes Aetiology Article Snippet: Membranes were blocked in blocking buffer (5% dry milk in PBS with 0.01% Tween) for one hour and incubated overnight at 4 °C with respective primary antibodies diluted in PBS with 5% dry milk and 0.01% Tween. .. The following antibodies and dilutions were used: anti-GLO1 (Sigma-Aldrich, St. Louis, MO, USA, SAB4200193, 1:4000) and anti-β-actin as a Article Title: Luteolin Induces microRNA-132 Expression and Modulates Neurite Outgrowth in PC12 Cells Article Snippet: Cell lysate (30 µg) was separated on 10% SDS-PAGE and transferred onto a PVDF membrane (PerkinElmer, Boston, MA, USA) at 25 volts overnight at 4°C. .. The membranes were blocked at 4°C in PBST blocking buffer (1% non-fat dried milk in PBS containing 0.1% Tween-20) for 8 h. Blots were incubated with the appropriate antibodies overnight at 4°C: anti-CREB (1∶1000), anti-phospho-CREB (Ser-133) (1∶1000), anti-ERK (1∶1000 ), anti-phospho-ERK (1∶1000 ) (Cell Signaling Technology, Inc.), and Electrophoresis:Article Title: Increased TIMP-3 expression alters the cellular secretome through dual inhibition of the metalloprotease ADAM10 and ligand-binding of the LRP-1 receptor Article Snippet: Supernatants from TIMP-3/HEK or HEK293 cells were loaded onto an acrylamide gel for electrophoresis. .. After electrophoretic separation, proteins were blotted onto a PVDF membrane using the Trans-Blot Turbo transfer system (Biorad) and detected by the following antibodies: anti-TIMP-3 (AB6000, Millipore), anti-TIMP-1 (generated as previously described ), anti-TIMP-2 (generated as previously described ), anti-SPARC (number 5031, Thermo Fisher), anti-MIF (clone FL-115, Santa Cruz) anti-APP (clone 22C11, Millipore), anti-MMP-1 (clone 2A7.2, Millipore) Article Title: Post-ischemic estradiol treatment reduced glial response and triggers distinct cortical and hippocampal signaling in a rat model of cerebral ischemia Article Snippet: After electrophoresis the proteins were electrotransferred onto nitrocellulose membranes (Whatman, Dassel, Germany) for 1.5 h at 100 mV using an electrophoretic transfer system (Mini-Trans-blot Electrophoretic Transfer Cell). .. Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), Modification:Article Title: Urinary exosomal expression of activator of G protein signaling 3 in polycystic kidney disease Article Snippet: Incubation:Article Title: Bioengineered NRF2-siRNA Is Effective to Interfere with NRF2 Pathways and Improve Chemosensitivity of Human Cancer Cells Article Snippet: The whole-cell proteins (35 μ g/lane) were resolved on a 10% SDS-PAGE gel and transferred to a polyvinylidene fluoride membrane. .. Membranes were incubated in 5% Blotting-Grade Blocker (catalog #170-6404; Bio-Rad) at room temperature for 2 hours, and then with primary antibodies, anti-NRF2 antibody (1:1000 dilution, catalog #ab62352; Abcam, Cambridge, MA), or Article Title: Generation and characterisation of a parkin-Pacrg knockout mouse line and a Pacrg knockout mouse line Article Snippet: PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 (A5441, Sigma-Aldrich). .. PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 ( Article Title: Deleterious Effect of Advanced CKD on Glyoxalase System Activity not Limited to Diabetes Aetiology Article Snippet: Membranes were blocked in blocking buffer (5% dry milk in PBS with 0.01% Tween) for one hour and incubated overnight at 4 °C with respective primary antibodies diluted in PBS with 5% dry milk and 0.01% Tween. .. The following antibodies and dilutions were used: anti-GLO1 (Sigma-Aldrich, St. Louis, MO, USA, SAB4200193, 1:4000) and anti-β-actin as a Article Title: Thyroxin Protects White Matter from Hypoxic-Ischemic Insult in the Immature Sprague–Dawley Rat Brain by Regulating Periventricular White Matter and Cortex BDNF and CREB Pathways Article Snippet: We detected proteins of interest with a chemiluminescence ECL system (GE Healthcare, Chicago, IL, USA) using secondary antibodies conjugated with horseradish peroxidase (Jackson ImmunoResearch, West Grove, PA, USA). .. The blots were stripped with buffer (2.5% SDS, 0.7% 2-mercaptoethanol, 62.5 mM Tris-HCl, pH 6.8) and incubated with the Article Title: Attenuation of autophagy flux by 6-shogaol sensitizes human liver cancer cells to TRAIL-induced apoptosis via p53 and ROS Article Snippet: Membranes were blocked with 5% non-fat dried milk at 25°C for 1 h, followed by incubation with primary antibodies overnight at 4°C. .. The Article Title: Post-ischemic estradiol treatment reduced glial response and triggers distinct cortical and hippocampal signaling in a rat model of cerebral ischemia Article Snippet: After electrophoresis the proteins were electrotransferred onto nitrocellulose membranes (Whatman, Dassel, Germany) for 1.5 h at 100 mV using an electrophoretic transfer system (Mini-Trans-blot Electrophoretic Transfer Cell). .. Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), Article Title: Myocardin Overexpression Is Sufficient for Promoting the Development of a Mature Smooth Muscle Cell-Like Phenotype from Human Embryonic Stem Cells Article Snippet: Proteins were extracted using lysis buffer (10 mM Tris-HCl, pH 7.6, 140 mM NaCl, 1 mM EDTA, 1% Triton X-100, protease and phosphatase inhibitor cocktail (Sigma)), and concentrations determined using a BCA Protein Assay Kit (Thermo Scientific). .. Samples were separated by SDS-PAGE and proteins transferred on to polyvinylidene difluoride membranes, blocked in 5% milk in Tris-buffered saline and 0.05% Tween 20, incubated with primary antibodies against myocardin (Sigma M8948; 1∶1000 dilution) and Article Title: Luteolin Induces microRNA-132 Expression and Modulates Neurite Outgrowth in PC12 Cells Article Snippet: Cell lysate (30 µg) was separated on 10% SDS-PAGE and transferred onto a PVDF membrane (PerkinElmer, Boston, MA, USA) at 25 volts overnight at 4°C. .. The membranes were blocked at 4°C in PBST blocking buffer (1% non-fat dried milk in PBS containing 0.1% Tween-20) for 8 h. Blots were incubated with the appropriate antibodies overnight at 4°C: anti-CREB (1∶1000), anti-phospho-CREB (Ser-133) (1∶1000), anti-ERK (1∶1000 ), anti-phospho-ERK (1∶1000 ) (Cell Signaling Technology, Inc.), and Stripping Membranes:Article Title: Generation and characterisation of a parkin-Pacrg knockout mouse line and a Pacrg knockout mouse line Article Snippet: PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 ( Significance Assay:Article Title: Increased TIMP-3 expression alters the cellular secretome through dual inhibition of the metalloprotease ADAM10 and ligand-binding of the LRP-1 receptor Article Snippet: After electrophoretic separation, proteins were blotted onto a PVDF membrane using the Trans-Blot Turbo transfer system (Biorad) and detected by the following antibodies: anti-TIMP-3 (AB6000, Millipore), anti-TIMP-1 (generated as previously described ), anti-TIMP-2 (generated as previously described ), anti-SPARC (number 5031, Thermo Fisher), anti-MIF (clone FL-115, Santa Cruz) anti-APP (clone 22C11, Millipore), anti-MMP-1 (clone 2A7.2, Millipore) Expressing:Article Title: Heterogeneity of p53-pathway Protein Expression in Chemosensitive Chronic Lymphocytic Leukemia: A Pilot Study Article Snippet: Paragraph title: Intracellular protein expression ... To control for protein loading, each membrane was re-probed with a mouse monoclonal antibody to BIA-KA:Article Title: Bioengineered NRF2-siRNA Is Effective to Interfere with NRF2 Pathways and Improve Chemosensitivity of Human Cancer Cells Article Snippet: Proteins were extracted from harvested cells using radioimmunoprecipitation lysis buffer supplemented with complete protease inhibitors, and protein concentrations were determined using the BCA Protein Assay Kit. .. Membranes were incubated in 5% Blotting-Grade Blocker (catalog #170-6404; Bio-Rad) at room temperature for 2 hours, and then with primary antibodies, anti-NRF2 antibody (1:1000 dilution, catalog #ab62352; Abcam, Cambridge, MA), or Article Title: Generation and characterisation of a parkin-Pacrg knockout mouse line and a Pacrg knockout mouse line Article Snippet: Protein concentration was determined using a BCA Protein Assay Kit (23225, Thermo Fisher Scientific). .. PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 ( Article Title: Deleterious Effect of Advanced CKD on Glyoxalase System Activity not Limited to Diabetes Aetiology Article Snippet: The following antibodies and dilutions were used: anti-GLO1 (Sigma-Aldrich, St. Louis, MO, USA, SAB4200193, 1:4000) and anti-β-actin as a reference protein (Sigma-Aldrich, St. Louis, MO, USA, A5441, 1:5000). .. The following antibodies and dilutions were used: anti-GLO1 (Sigma-Aldrich, St. Louis, MO, USA, SAB4200193, 1:4000) and anti-β-actin as a Article Title: Thyroxin Protects White Matter from Hypoxic-Ischemic Insult in the Immature Sprague–Dawley Rat Brain by Regulating Periventricular White Matter and Cortex BDNF and CREB Pathways Article Snippet: Protein concentrations of supernatant were determined using a BCA protein-assay kit (Pierce Kit #23227, Thermo Scientific, Waltham, MA, USA) with bovine-serum albumin to plot a standard curve. .. The blots were stripped with buffer (2.5% SDS, 0.7% 2-mercaptoethanol, 62.5 mM Tris-HCl, pH 6.8) and incubated with the Article Title: Attenuation of autophagy flux by 6-shogaol sensitizes human liver cancer cells to TRAIL-induced apoptosis via p53 and ROS Article Snippet: The protein concentration was determined using the Pierce BCA Protein Assay kit (Thermo Fisher Scientific, Inc.). .. The Article Title: Biochemical alterations in inflammatory reactive chondrocytes: evidence for intercellular network communication Article Snippet: Protein concentration was determined using the Pierce BCA protein assay kit (Life Technologies), according to the manufacturer’s instructions. .. The membranes were probed with the following primary antibodies; polyclonal rabbit anti-TLR-4 (M-300) (sc-30002, Santa Cruz), polyclonal rabbit anti-GLT-1 (pab0037, Covalab), polyclonal rabbit anti-GLAST (pab0036-P, Covalab), polyclonal rabbit anti-connexin 4371–0700, Life technologies), monoclonal mouse anti-Na+/K+-ATPase (A276, Sigma-Aldrich), and Article Title: Myocardin Overexpression Is Sufficient for Promoting the Development of a Mature Smooth Muscle Cell-Like Phenotype from Human Embryonic Stem Cells Article Snippet: Proteins were extracted using lysis buffer (10 mM Tris-HCl, pH 7.6, 140 mM NaCl, 1 mM EDTA, 1% Triton X-100, protease and phosphatase inhibitor cocktail (Sigma)), and concentrations determined using a BCA Protein Assay Kit (Thermo Scientific). .. Samples were separated by SDS-PAGE and proteins transferred on to polyvinylidene difluoride membranes, blocked in 5% milk in Tris-buffered saline and 0.05% Tween 20, incubated with primary antibodies against myocardin (Sigma M8948; 1∶1000 dilution) and Acrylamide Gel Assay:Article Title: Increased TIMP-3 expression alters the cellular secretome through dual inhibition of the metalloprotease ADAM10 and ligand-binding of the LRP-1 receptor Article Snippet: Supernatants from TIMP-3/HEK or HEK293 cells were loaded onto an acrylamide gel for electrophoresis. .. After electrophoretic separation, proteins were blotted onto a PVDF membrane using the Trans-Blot Turbo transfer system (Biorad) and detected by the following antibodies: anti-TIMP-3 (AB6000, Millipore), anti-TIMP-1 (generated as previously described ), anti-TIMP-2 (generated as previously described ), anti-SPARC (number 5031, Thermo Fisher), anti-MIF (clone FL-115, Santa Cruz) anti-APP (clone 22C11, Millipore), anti-MMP-1 (clone 2A7.2, Millipore) Western Blot:Article Title: P2X7 signaling promotes microsphere embolism-triggered microglia activation by maintaining elevation of Fas ligand Article Snippet: Paragraph title: Western blotting ... Membranes were probed overnight at 4°C with antibodies against Fas (sc-736), FasL (sc-6237), P2X7 (sc-25698), High mobility group box 1 (HMGB1, sc-26351), caspase-8 (sc-7890) (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA); CD3 (Abcam Cambridge, UK, Catalog no: ab5690); FADD (Epitomics, Inc., Burlingame, CA, USA, Catalog no: 2988–1); and Article Title: Bioengineered NRF2-siRNA Is Effective to Interfere with NRF2 Pathways and Improve Chemosensitivity of Human Cancer Cells Article Snippet: Paragraph title: Protein Isolation and Western Blots. ... Membranes were incubated in 5% Blotting-Grade Blocker (catalog #170-6404; Bio-Rad) at room temperature for 2 hours, and then with primary antibodies, anti-NRF2 antibody (1:1000 dilution, catalog #ab62352; Abcam, Cambridge, MA), or Article Title: Generation and characterisation of a parkin-Pacrg knockout mouse line and a Pacrg knockout mouse line Article Snippet: Paragraph title: Western blot analysis ... PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 ( Article Title: Urinary exosomal expression of activator of G protein signaling 3 in polycystic kidney disease Article Snippet: AGS3 protein detection was determined using standard Western blot techniques [ , , ]. .. Article Title: Deleterious Effect of Advanced CKD on Glyoxalase System Activity not Limited to Diabetes Aetiology Article Snippet: Paragraph title: 4.5. Protein Isolation and Western Blotting ... The following antibodies and dilutions were used: anti-GLO1 (Sigma-Aldrich, St. Louis, MO, USA, SAB4200193, 1:4000) and anti-β-actin as a Article Title: Thyroxin Protects White Matter from Hypoxic-Ischemic Insult in the Immature Sprague–Dawley Rat Brain by Regulating Periventricular White Matter and Cortex BDNF and CREB Pathways Article Snippet: Paragraph title: 4.8. Western Blotting ... The blots were stripped with buffer (2.5% SDS, 0.7% 2-mercaptoethanol, 62.5 mM Tris-HCl, pH 6.8) and incubated with the Article Title: Attenuation of autophagy flux by 6-shogaol sensitizes human liver cancer cells to TRAIL-induced apoptosis via p53 and ROS Article Snippet: Paragraph title: Western blot assay ... The Article Title: Biochemical alterations in inflammatory reactive chondrocytes: evidence for intercellular network communication Article Snippet: Paragraph title: 2.1.6. Western blot analysis ... The membranes were probed with the following primary antibodies; polyclonal rabbit anti-TLR-4 (M-300) (sc-30002, Santa Cruz), polyclonal rabbit anti-GLT-1 (pab0037, Covalab), polyclonal rabbit anti-GLAST (pab0036-P, Covalab), polyclonal rabbit anti-connexin 4371–0700, Life technologies), monoclonal mouse anti-Na+/K+-ATPase (A276, Sigma-Aldrich), and Article Title: Transcriptional coactivator PGC-1α contains a novel CBP80-binding motif that orchestrates efficient target gene expression Article Snippet: Paragraph title: Western blotting ... Proteins were detected using the following antibodies: anti-PGC-1β (Bethyl Laboratories, A302-273A), anti-PGC-1α (Novus, NBP1-04676), anti-Flag (Sigma-Aldrich, A8592), Article Title: Post-ischemic estradiol treatment reduced glial response and triggers distinct cortical and hippocampal signaling in a rat model of cerebral ischemia Article Snippet: Paragraph title: Western blotting ... Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), Article Title: Myocardin Overexpression Is Sufficient for Promoting the Development of a Mature Smooth Muscle Cell-Like Phenotype from Human Embryonic Stem Cells Article Snippet: Paragraph title: Western Blot Analysis ... Samples were separated by SDS-PAGE and proteins transferred on to polyvinylidene difluoride membranes, blocked in 5% milk in Tris-buffered saline and 0.05% Tween 20, incubated with primary antibodies against myocardin (Sigma M8948; 1∶1000 dilution) and Article Title: Expression of Extracellular Superoxide Dismutase Protein in Diabetes Article Snippet: Paragraph title: Western blot analysis ... The membranes were stripped and reblotted with Article Title: LC3B is not recruited along with the autophagy elongation complex (ATG5-12/16L1) at HCV replication site and is dispensable for viral replication Article Snippet: The Flag-tagged ATG12 (pATG12) and its dominant-negative derivative pATG12ΔG140 (ATG12-DN) constructs were kindly provided by Dr. Adi Kimchi (Israel) [ ]. .. Rabbit polyclonal anti-LC3, rabbit polyclonal anti-ATG5 (used for western blot), mouse monoclonal anti-Flag, and Article Title: Herpes Simplex Virus Type 1 Single Strand DNA Binding Protein and Helicase/Primase Complex Disable Cellular ATR Signaling Article Snippet: Paragraph title: Western blot analysis ... Primary antibodies used include polyclonal rabbit anti-ATRIP 403 (rATRIP 403) (1∶3,000) , monoclonal mouse anti-ICP4 (1∶10,000; US Biologics), Article Title: ENaC-mediated alveolar fluid clearance and lung fluid balance depend on the channel-activating protease 1 Article Snippet: Paragraph title: Distal lung protein extraction and Western blot analysis ... Samples of protein extracts (100–200 µg/sample, the amount necessary to detect minor bands of ENaC proteins) were separated by SDS-PAGE on 8% acrylamide gels, electrically transferred to nitrocellulose paper, and subsequently probed for ENaC subunits and β-actin by using previously characterized rabbit polyclonal anti rat α-, β- and γ-ENaC antibodies (dilution 1:2000) (Duc et al, ), and Article Title: Luteolin Induces microRNA-132 Expression and Modulates Neurite Outgrowth in PC12 Cells Article Snippet: Paragraph title: Western Blotting Analysis of CREB, Phospho-CREB, ERK, Phospho-ERK and β-actin Proteins ... The membranes were blocked at 4°C in PBST blocking buffer (1% non-fat dried milk in PBS containing 0.1% Tween-20) for 8 h. Blots were incubated with the appropriate antibodies overnight at 4°C: anti-CREB (1∶1000), anti-phospho-CREB (Ser-133) (1∶1000), anti-ERK (1∶1000 ), anti-phospho-ERK (1∶1000 ) (Cell Signaling Technology, Inc.), and Protease Inhibitor:Article Title: Biochemical alterations in inflammatory reactive chondrocytes: evidence for intercellular network communication Article Snippet: Briefly, protein extracts were prepared by cell lysis in RIPA buffer (Sigma-Aldrich) supplemented with a mammalian protease inhibitor cocktail (Sigma-Aldrich). .. The membranes were probed with the following primary antibodies; polyclonal rabbit anti-TLR-4 (M-300) (sc-30002, Santa Cruz), polyclonal rabbit anti-GLT-1 (pab0037, Covalab), polyclonal rabbit anti-GLAST (pab0036-P, Covalab), polyclonal rabbit anti-connexin 4371–0700, Life technologies), monoclonal mouse anti-Na+/K+-ATPase (A276, Sigma-Aldrich), and Article Title: Herpes Simplex Virus Type 1 Single Strand DNA Binding Protein and Helicase/Primase Complex Disable Cellular ATR Signaling Article Snippet: Primary antibodies used include polyclonal rabbit anti-ATRIP 403 (rATRIP 403) (1∶3,000) , monoclonal mouse anti-ICP4 (1∶10,000; US Biologics), monoclonal mouse anti-β-actin (1∶15,000; Sigma), polyclonal goat anti-ATR N19 (1∶1,000; Santa Cruz), monoclonal mouse anti-Chk1 (1∶1,000; Santa Cruz), monoclonal mouse anti-HA (F7) (1∶3,000; Santa Cruz), monoclonal mouse anti-RPA32 (9H8) (1∶1,000; Genetex), monoclonal rabbit anti-phospho-Chk1 S345 (1∶5,000; Cell Signaling), polyclonal rabbit anti-phospho-RPA S33 (1∶3,000; Bethyl), and polyclonal rabbit anti-phospho-RPA S4/S8 (1∶3,000; Bethyl). .. Primary antibodies used include polyclonal rabbit anti-ATRIP 403 (rATRIP 403) (1∶3,000) , monoclonal mouse anti-ICP4 (1∶10,000; US Biologics), Cell Culture:Article Title: Heterogeneity of p53-pathway Protein Expression in Chemosensitive Chronic Lymphocytic Leukemia: A Pilot Study Article Snippet: Proteins from normal blood MNC cultured with cisplatin and phytohemagglutinin (PHA), and untreated MNC probed with antibodies on the same membrane served as positive and negative controls respectively. .. To control for protein loading, each membrane was re-probed with a mouse monoclonal antibody to Generated:Article Title: Increased TIMP-3 expression alters the cellular secretome through dual inhibition of the metalloprotease ADAM10 and ligand-binding of the LRP-1 receptor Article Snippet: Supernatants from TIMP-3/HEK or HEK293 cells were loaded onto an acrylamide gel for electrophoresis. .. After electrophoretic separation, proteins were blotted onto a PVDF membrane using the Trans-Blot Turbo transfer system (Biorad) and detected by the following antibodies: anti-TIMP-3 (AB6000, Millipore), anti-TIMP-1 (generated as previously described ), anti-TIMP-2 (generated as previously described ), anti-SPARC (number 5031, Thermo Fisher), anti-MIF (clone FL-115, Santa Cruz) anti-APP (clone 22C11, Millipore), anti-MMP-1 (clone 2A7.2, Millipore) other:Article Title: Insulin-like growth factor receptor signaling in breast tumor epithelium protects cells from endoplasmic reticulum stress and regulates the tumor microenvironment Article Snippet: Imaging:Article Title: Bioengineered NRF2-siRNA Is Effective to Interfere with NRF2 Pathways and Improve Chemosensitivity of Human Cancer Cells Article Snippet: Membranes were incubated in 5% Blotting-Grade Blocker (catalog #170-6404; Bio-Rad) at room temperature for 2 hours, and then with primary antibodies, anti-NRF2 antibody (1:1000 dilution, catalog #ab62352; Abcam, Cambridge, MA), or Protein Concentration:Article Title: Generation and characterisation of a parkin-Pacrg knockout mouse line and a Pacrg knockout mouse line Article Snippet: Protein concentration was determined using a BCA Protein Assay Kit (23225, Thermo Fisher Scientific). .. PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 ( Article Title: Attenuation of autophagy flux by 6-shogaol sensitizes human liver cancer cells to TRAIL-induced apoptosis via p53 and ROS Article Snippet: The protein concentration was determined using the Pierce BCA Protein Assay kit (Thermo Fisher Scientific, Inc.). .. The Article Title: Biochemical alterations in inflammatory reactive chondrocytes: evidence for intercellular network communication Article Snippet: Protein concentration was determined using the Pierce BCA protein assay kit (Life Technologies), according to the manufacturer’s instructions. .. The membranes were probed with the following primary antibodies; polyclonal rabbit anti-TLR-4 (M-300) (sc-30002, Santa Cruz), polyclonal rabbit anti-GLT-1 (pab0037, Covalab), polyclonal rabbit anti-GLAST (pab0036-P, Covalab), polyclonal rabbit anti-connexin 4371–0700, Life technologies), monoclonal mouse anti-Na+/K+-ATPase (A276, Sigma-Aldrich), and Article Title: Post-ischemic estradiol treatment reduced glial response and triggers distinct cortical and hippocampal signaling in a rat model of cerebral ischemia Article Snippet: The protein concentration of each sample was determined using the DC Protein Assay kit (BioRad, Hercules, CA, USA) according to the manufacturer’s instructions. .. Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), Article Title: Luteolin Induces microRNA-132 Expression and Modulates Neurite Outgrowth in PC12 Cells Article Snippet: The protein concentration was measured by the Bradford method (Bio-Rad Laboratories, Hercules, CA, USA) using bovine serum albumin as the standard. .. The membranes were blocked at 4°C in PBST blocking buffer (1% non-fat dried milk in PBS containing 0.1% Tween-20) for 8 h. Blots were incubated with the appropriate antibodies overnight at 4°C: anti-CREB (1∶1000), anti-phospho-CREB (Ser-133) (1∶1000), anti-ERK (1∶1000 ), anti-phospho-ERK (1∶1000 ) (Cell Signaling Technology, Inc.), and Binding Assay:Article Title: Post-ischemic estradiol treatment reduced glial response and triggers distinct cortical and hippocampal signaling in a rat model of cerebral ischemia Article Snippet: Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), Molecular Weight:Article Title: Thyroxin Protects White Matter from Hypoxic-Ischemic Insult in the Immature Sprague–Dawley Rat Brain by Regulating Periventricular White Matter and Cortex BDNF and CREB Pathways Article Snippet: After denaturing in Laemmli buffer (catalog #161-0737, Bio-Rad, Hercules, CA, USA), equal amounts of protein (10–20 μg) were loaded onto 4–15% or 4–20% gradient precast gels (Bio-Rad), depending on the molecular weight of the target protein. .. The blots were stripped with buffer (2.5% SDS, 0.7% 2-mercaptoethanol, 62.5 mM Tris-HCl, pH 6.8) and incubated with the DC Protein Assay:Article Title: Post-ischemic estradiol treatment reduced glial response and triggers distinct cortical and hippocampal signaling in a rat model of cerebral ischemia Article Snippet: The protein concentration of each sample was determined using the DC Protein Assay kit (BioRad, Hercules, CA, USA) according to the manufacturer’s instructions. .. Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), Isolation:Article Title: P2X7 signaling promotes microsphere embolism-triggered microglia activation by maintaining elevation of Fas ligand Article Snippet: Proteins were extracted, and the soluble cytosolic fraction was isolated according to our previous study [ ]. .. Membranes were probed overnight at 4°C with antibodies against Fas (sc-736), FasL (sc-6237), P2X7 (sc-25698), High mobility group box 1 (HMGB1, sc-26351), caspase-8 (sc-7890) (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA); CD3 (Abcam Cambridge, UK, Catalog no: ab5690); FADD (Epitomics, Inc., Burlingame, CA, USA, Catalog no: 2988–1); and Article Title: Bioengineered NRF2-siRNA Is Effective to Interfere with NRF2 Pathways and Improve Chemosensitivity of Human Cancer Cells Article Snippet: Paragraph title: Protein Isolation and Western Blots. ... Membranes were incubated in 5% Blotting-Grade Blocker (catalog #170-6404; Bio-Rad) at room temperature for 2 hours, and then with primary antibodies, anti-NRF2 antibody (1:1000 dilution, catalog #ab62352; Abcam, Cambridge, MA), or Article Title: Urinary exosomal expression of activator of G protein signaling 3 in polycystic kidney disease Article Snippet: SD and PCK rat kidney lysates at 8, 16, and 24 weeks of age, and human and rat urine exosome protein lysates were isolated using 1X RIPA buffer containing protease (Roche) and phosphatase inhibitors (Pierce, Rockford, IL). .. Article Title: Deleterious Effect of Advanced CKD on Glyoxalase System Activity not Limited to Diabetes Aetiology Article Snippet: Paragraph title: 4.5. Protein Isolation and Western Blotting ... The following antibodies and dilutions were used: anti-GLO1 (Sigma-Aldrich, St. Louis, MO, USA, SAB4200193, 1:4000) and anti-β-actin as a Protein Extraction:Article Title: ENaC-mediated alveolar fluid clearance and lung fluid balance depend on the channel-activating protease 1 Article Snippet: Paragraph title: Distal lung protein extraction and Western blot analysis ... Samples of protein extracts (100–200 µg/sample, the amount necessary to detect minor bands of ENaC proteins) were separated by SDS-PAGE on 8% acrylamide gels, electrically transferred to nitrocellulose paper, and subsequently probed for ENaC subunits and β-actin by using previously characterized rabbit polyclonal anti rat α-, β- and γ-ENaC antibodies (dilution 1:2000) (Duc et al, ), and Construct:Article Title: LC3B is not recruited along with the autophagy elongation complex (ATG5-12/16L1) at HCV replication site and is dispensable for viral replication Article Snippet: The Flag-tagged ATG12 (pATG12) and its dominant-negative derivative pATG12ΔG140 (ATG12-DN) constructs were kindly provided by Dr. Adi Kimchi (Israel) [ ]. .. Rabbit polyclonal anti-LC3, rabbit polyclonal anti-ATG5 (used for western blot), mouse monoclonal anti-Flag, and Polyacrylamide Gel Electrophoresis:Article Title: P2X7 signaling promotes microsphere embolism-triggered microglia activation by maintaining elevation of Fas ligand Article Snippet: Samples containing equivalent amounts of protein were loaded onto 10% to 15% acrylamide denaturing gels (sodium dodecyl sulfate polyacrylamide gel electrophoresis; SDS-PAGE). .. Membranes were probed overnight at 4°C with antibodies against Fas (sc-736), FasL (sc-6237), P2X7 (sc-25698), High mobility group box 1 (HMGB1, sc-26351), caspase-8 (sc-7890) (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA); CD3 (Abcam Cambridge, UK, Catalog no: ab5690); FADD (Epitomics, Inc., Burlingame, CA, USA, Catalog no: 2988–1); and Staining:Article Title: Biochemical alterations in inflammatory reactive chondrocytes: evidence for intercellular network communication Article Snippet: The membranes were probed with the following primary antibodies; polyclonal rabbit anti-TLR-4 (M-300) (sc-30002, Santa Cruz), polyclonal rabbit anti-GLT-1 (pab0037, Covalab), polyclonal rabbit anti-GLAST (pab0036-P, Covalab), polyclonal rabbit anti-connexin 4371–0700, Life technologies), monoclonal mouse anti-Na+/K+-ATPase (A276, Sigma-Aldrich), and mouse monoclonal anti-β-actin (A5441, Sigma-Aldrich). .. The membranes were probed with the following primary antibodies; polyclonal rabbit anti-TLR-4 (M-300) (sc-30002, Santa Cruz), polyclonal rabbit anti-GLT-1 (pab0037, Covalab), polyclonal rabbit anti-GLAST (pab0036-P, Covalab), polyclonal rabbit anti-connexin 4371–0700, Life technologies), monoclonal mouse anti-Na+/K+-ATPase (A276, Sigma-Aldrich), and SDS Page:Article Title: P2X7 signaling promotes microsphere embolism-triggered microglia activation by maintaining elevation of Fas ligand Article Snippet: Samples containing equivalent amounts of protein were loaded onto 10% to 15% acrylamide denaturing gels (sodium dodecyl sulfate polyacrylamide gel electrophoresis; SDS-PAGE). .. Membranes were probed overnight at 4°C with antibodies against Fas (sc-736), FasL (sc-6237), P2X7 (sc-25698), High mobility group box 1 (HMGB1, sc-26351), caspase-8 (sc-7890) (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA); CD3 (Abcam Cambridge, UK, Catalog no: ab5690); FADD (Epitomics, Inc., Burlingame, CA, USA, Catalog no: 2988–1); and Article Title: Bioengineered NRF2-siRNA Is Effective to Interfere with NRF2 Pathways and Improve Chemosensitivity of Human Cancer Cells Article Snippet: The whole-cell proteins (35 μ g/lane) were resolved on a 10% SDS-PAGE gel and transferred to a polyvinylidene fluoride membrane. .. Membranes were incubated in 5% Blotting-Grade Blocker (catalog #170-6404; Bio-Rad) at room temperature for 2 hours, and then with primary antibodies, anti-NRF2 antibody (1:1000 dilution, catalog #ab62352; Abcam, Cambridge, MA), or Article Title: Generation and characterisation of a parkin-Pacrg knockout mouse line and a Pacrg knockout mouse line Article Snippet: Approximately 50 μg of lysate was fractioned in 15% SDS-PAGE gels that were subsequently transferred to PVDF Immobilon-P membranes (IPVH00010, Millipore-Merck). .. PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 ( Article Title: Deleterious Effect of Advanced CKD on Glyoxalase System Activity not Limited to Diabetes Aetiology Article Snippet: Twenty micrograms (20 μg) of protein lysates were separated in 1.5 mm thick 12% SDS-PAGE gels. .. The following antibodies and dilutions were used: anti-GLO1 (Sigma-Aldrich, St. Louis, MO, USA, SAB4200193, 1:4000) and anti-β-actin as a Article Title: Attenuation of autophagy flux by 6-shogaol sensitizes human liver cancer cells to TRAIL-induced apoptosis via p53 and ROS Article Snippet: Proteins (30 µ g) were separated on 10% SDS-PAGE gels and blotted onto polyvinylidene fluoride membranes. .. The Article Title: Post-ischemic estradiol treatment reduced glial response and triggers distinct cortical and hippocampal signaling in a rat model of cerebral ischemia Article Snippet: The proteins were resolved by SDS-PAGE (8%) using a Mini-Protean system (Bio-Rad) and depending on the epitope to be analyzed, 30–50 μg of total protein was loaded into each lane in loading buffer containing: 0.062 M Tris (pH 6.8), 10% glycerol, 5% β-mercaptoethanol, 7.5 mM EDTA, 2% SDS, and 0.002% bromophenol blue. .. Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), Article Title: Myocardin Overexpression Is Sufficient for Promoting the Development of a Mature Smooth Muscle Cell-Like Phenotype from Human Embryonic Stem Cells Article Snippet: Proteins were extracted using lysis buffer (10 mM Tris-HCl, pH 7.6, 140 mM NaCl, 1 mM EDTA, 1% Triton X-100, protease and phosphatase inhibitor cocktail (Sigma)), and concentrations determined using a BCA Protein Assay Kit (Thermo Scientific). .. Samples were separated by SDS-PAGE and proteins transferred on to polyvinylidene difluoride membranes, blocked in 5% milk in Tris-buffered saline and 0.05% Tween 20, incubated with primary antibodies against myocardin (Sigma M8948; 1∶1000 dilution) and Article Title: Expression of Extracellular Superoxide Dismutase Protein in Diabetes Article Snippet: Proteins from tissue were separated by SDS-PAGE using NuPAGE 4% to 12% bis-Tris gels (Invitrogen, Carlsbad, CA, USA, NP0335Box) and then transferred to Immobilon-P membrane. .. The membranes were stripped and reblotted with Article Title: Herpes Simplex Virus Type 1 Single Strand DNA Binding Protein and Helicase/Primase Complex Disable Cellular ATR Signaling Article Snippet: Proteins were resolved by SDS-PAGE and transferred to PVDF membranes. .. Primary antibodies used include polyclonal rabbit anti-ATRIP 403 (rATRIP 403) (1∶3,000) , monoclonal mouse anti-ICP4 (1∶10,000; US Biologics), Article Title: ENaC-mediated alveolar fluid clearance and lung fluid balance depend on the channel-activating protease 1 Article Snippet: Lungs were removed from thorax and homogenized for 3 min in ice-cold lysis RIPA buffer (pH 8) containing 20 mM Tris, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 0.5% deoxycholate and protease inhibitors. .. Samples of protein extracts (100–200 µg/sample, the amount necessary to detect minor bands of ENaC proteins) were separated by SDS-PAGE on 8% acrylamide gels, electrically transferred to nitrocellulose paper, and subsequently probed for ENaC subunits and β-actin by using previously characterized rabbit polyclonal anti rat α-, β- and γ-ENaC antibodies (dilution 1:2000) (Duc et al, ), and Article Title: Luteolin Induces microRNA-132 Expression and Modulates Neurite Outgrowth in PC12 Cells Article Snippet: Cell lysate (30 µg) was separated on 10% SDS-PAGE and transferred onto a PVDF membrane (PerkinElmer, Boston, MA, USA) at 25 volts overnight at 4°C. .. The membranes were blocked at 4°C in PBST blocking buffer (1% non-fat dried milk in PBS containing 0.1% Tween-20) for 8 h. Blots were incubated with the appropriate antibodies overnight at 4°C: anti-CREB (1∶1000), anti-phospho-CREB (Ser-133) (1∶1000), anti-ERK (1∶1000 ), anti-phospho-ERK (1∶1000 ) (Cell Signaling Technology, Inc.), and Plasmid Preparation:Article Title: LC3B is not recruited along with the autophagy elongation complex (ATG5-12/16L1) at HCV replication site and is dispensable for viral replication Article Snippet: hATG5 and hATG16L1 sequences were cloned into peGFP-C1 plasmid (Clontech) to form pGFP-ATG5 and pGFP-ATG16L1, respectively. .. Rabbit polyclonal anti-LC3, rabbit polyclonal anti-ATG5 (used for western blot), mouse monoclonal anti-Flag, and Software:Article Title: Bioengineered NRF2-siRNA Is Effective to Interfere with NRF2 Pathways and Improve Chemosensitivity of Human Cancer Cells Article Snippet: Membranes were incubated in 5% Blotting-Grade Blocker (catalog #170-6404; Bio-Rad) at room temperature for 2 hours, and then with primary antibodies, anti-NRF2 antibody (1:1000 dilution, catalog #ab62352; Abcam, Cambridge, MA), or Article Title: Generation and characterisation of a parkin-Pacrg knockout mouse line and a Pacrg knockout mouse line Article Snippet: PACRG was detected using rabbit anti-PACRG clone MC1290 diluted 1:1000 . β-Actin was used as a loading control and was detected by mouse anti-β-Actin ascites clone AC-15 diluted 1:10000 ( Article Title: Urinary exosomal expression of activator of G protein signaling 3 in polycystic kidney disease Article Snippet: Article Title: Deleterious Effect of Advanced CKD on Glyoxalase System Activity not Limited to Diabetes Aetiology Article Snippet: The following antibodies and dilutions were used: anti-GLO1 (Sigma-Aldrich, St. Louis, MO, USA, SAB4200193, 1:4000) and anti-β-actin as a Article Title: Biochemical alterations in inflammatory reactive chondrocytes: evidence for intercellular network communication Article Snippet: The membranes were probed with the following primary antibodies; polyclonal rabbit anti-TLR-4 (M-300) (sc-30002, Santa Cruz), polyclonal rabbit anti-GLT-1 (pab0037, Covalab), polyclonal rabbit anti-GLAST (pab0036-P, Covalab), polyclonal rabbit anti-connexin 4371–0700, Life technologies), monoclonal mouse anti-Na+/K+-ATPase (A276, Sigma-Aldrich), and Article Title: Post-ischemic estradiol treatment reduced glial response and triggers distinct cortical and hippocampal signaling in a rat model of cerebral ischemia Article Snippet: Subsequently, the membranes were blocked for 1 h with 5% non-fat powdered milk in phosphate buffered saline (PBS) containing 0.1% Tween-20 (PBS-T) and then they were incubated overnight at 4 °C with the appropriate primary antibody: Akt (1:1,000; Cell Signaling no. 9272); phospho-AktSer473 (1:500; Cell Signaling no. 9271), phospho-AktThr308 (1:1,000; clone C31E5E, Cell Signaling no. 2965), β-catenin (1:800; BD Transduction Laboratories no. 610153), β-tubulin (1:1,000; clone TUB 2.1, Sigma no. T4026), Article Title: ENaC-mediated alveolar fluid clearance and lung fluid balance depend on the channel-activating protease 1 Article Snippet: Samples of protein extracts (100–200 µg/sample, the amount necessary to detect minor bands of ENaC proteins) were separated by SDS-PAGE on 8% acrylamide gels, electrically transferred to nitrocellulose paper, and subsequently probed for ENaC subunits and β-actin by using previously characterized rabbit polyclonal anti rat α-, β- and γ-ENaC antibodies (dilution 1:2000) (Duc et al, ), and Dominant Negative Mutation:Article Title: LC3B is not recruited along with the autophagy elongation complex (ATG5-12/16L1) at HCV replication site and is dispensable for viral replication Article Snippet: The Flag-tagged ATG12 (pATG12) and its dominant-negative derivative pATG12ΔG140 (ATG12-DN) constructs were kindly provided by Dr. Adi Kimchi (Israel) [ ]. .. Rabbit polyclonal anti-LC3, rabbit polyclonal anti-ATG5 (used for western blot), mouse monoclonal anti-Flag, and In Vitro:Article Title: Heterogeneity of p53-pathway Protein Expression in Chemosensitive Chronic Lymphocytic Leukemia: A Pilot Study Article Snippet: To control for protein loading, each membrane was re-probed with a mouse monoclonal antibody to Radio Immunoprecipitation:Article Title: Attenuation of autophagy flux by 6-shogaol sensitizes human liver cancer cells to TRAIL-induced apoptosis via p53 and ROS Article Snippet: Briefly, radioimmunoprecipitation assay buffer (Qiagen, Inc., Valencia, CA, USA) was used to extract total proteins from Huh7 cells. .. The Concentration Assay:Article Title: Deleterious Effect of Advanced CKD on Glyoxalase System Activity not Limited to Diabetes Aetiology Article Snippet: The following antibodies and dilutions were used: anti-GLO1 (Sigma-Aldrich, St. Louis, MO, USA, SAB4200193, 1:4000) and anti-β-actin as a reference protein (Sigma-Aldrich, St. Louis, MO, USA, A5441, 1:5000). .. The following antibodies and dilutions were used: anti-GLO1 (Sigma-Aldrich, St. Louis, MO, USA, SAB4200193, 1:4000) and anti-β-actin as a Lysis:Article Title: Bioengineered NRF2-siRNA Is Effective to Interfere with NRF2 Pathways and Improve Chemosensitivity of Human Cancer Cells Article Snippet: Proteins were extracted from harvested cells using radioimmunoprecipitation lysis buffer supplemented with complete protease inhibitors, and protein concentrations were determined using the BCA Protein Assay Kit. .. Membranes were incubated in 5% Blotting-Grade Blocker (catalog #170-6404; Bio-Rad) at room temperature for 2 hours, and then with primary antibodies, anti-NRF2 antibody (1:1000 dilution, catalog #ab62352; Abcam, Cambridge, MA), or Article Title: Biochemical alterations in inflammatory reactive chondrocytes: evidence for intercellular network communication Article Snippet: Briefly, protein extracts were prepared by cell lysis in RIPA buffer (Sigma-Aldrich) supplemented with a mammalian protease inhibitor cocktail (Sigma-Aldrich). .. The membranes were probed with the following primary antibodies; polyclonal rabbit anti-TLR-4 (M-300) (sc-30002, Santa Cruz), polyclonal rabbit anti-GLT-1 (pab0037, Covalab), polyclonal rabbit anti-GLAST (pab0036-P, Covalab), polyclonal rabbit anti-connexin 4371–0700, Life technologies), monoclonal mouse anti-Na+/K+-ATPase (A276, Sigma-Aldrich), and Article Title: Myocardin Overexpression Is Sufficient for Promoting the Development of a Mature Smooth Muscle Cell-Like Phenotype from Human Embryonic Stem Cells Article Snippet: Proteins were extracted using lysis buffer (10 mM Tris-HCl, pH 7.6, 140 mM NaCl, 1 mM EDTA, 1% Triton X-100, protease and phosphatase inhibitor cocktail (Sigma)), and concentrations determined using a BCA Protein Assay Kit (Thermo Scientific). .. Samples were separated by SDS-PAGE and proteins transferred on to polyvinylidene difluoride membranes, blocked in 5% milk in Tris-buffered saline and 0.05% Tween 20, incubated with primary antibodies against myocardin (Sigma M8948; 1∶1000 dilution) and Article Title: ENaC-mediated alveolar fluid clearance and lung fluid balance depend on the channel-activating protease 1 Article Snippet: Lungs were removed from thorax and homogenized for 3 min in ice-cold lysis RIPA buffer (pH 8) containing 20 mM Tris, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 0.5% deoxycholate and protease inhibitors. .. Samples of protein extracts (100–200 µg/sample, the amount necessary to detect minor bands of ENaC proteins) were separated by SDS-PAGE on 8% acrylamide gels, electrically transferred to nitrocellulose paper, and subsequently probed for ENaC subunits and β-actin by using previously characterized rabbit polyclonal anti rat α-, β- and γ-ENaC antibodies (dilution 1:2000) (Duc et al, ), and T-Test:Article Title: Increased TIMP-3 expression alters the cellular secretome through dual inhibition of the metalloprotease ADAM10 and ligand-binding of the LRP-1 receptor Article Snippet: After electrophoretic separation, proteins were blotted onto a PVDF membrane using the Trans-Blot Turbo transfer system (Biorad) and detected by the following antibodies: anti-TIMP-3 (AB6000, Millipore), anti-TIMP-1 (generated as previously described ), anti-TIMP-2 (generated as previously described ), anti-SPARC (number 5031, Thermo Fisher), anti-MIF (clone FL-115, Santa Cruz) anti-APP (clone 22C11, Millipore), anti-MMP-1 (clone 2A7.2, Millipore) Activity Assay:Article Title: Reduced PKC α Activity Induces Senescent Phenotype in Erythrocytes Article Snippet: PMA (P 8139), 4α -phorbol 12,13-didecanoate (4α PDD) (P 8014), phenyl methyl sulfonyl fluoride (PMSF) (P 7626), anti-band 3 N-terminus monoclonal antibody (B 9277), and |