monoclonal anti beta actin  (Millipore)


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    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 α actins skeletal cardiac smooth one β actin β non muscle and two γ actins γ smooth muscle and γ non muscle ACTB actin β gene codes for β actin It is a cytoskeletal housekeeping protein It is present in the cytoplasm The ACTB gene is located on human chromosome 7p22 1 Monoclonal Anti β Actin mouse IgG2a isotype is derived from the AC 74 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from an immunized mouse
    Catalog Number:
    a5316
    Price:
    None
    Applications:
    Western blot analysis of MDCK cell lysates were performed using monoclonal anti-actin antibody as a primary antibody.
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    Structured Review

    Millipore monoclonal anti beta actin
    Monoclonal Anti beta Actin antibody
    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 α actins skeletal cardiac smooth one β actin β non muscle and two γ actins γ smooth muscle and γ non muscle ACTB actin β gene codes for β actin It is a cytoskeletal housekeeping protein It is present in the cytoplasm The ACTB gene is located on human chromosome 7p22 1 Monoclonal Anti β Actin mouse IgG2a isotype is derived from the AC 74 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from an immunized mouse
    https://www.bioz.com/result/monoclonal anti beta actin/product/Millipore
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti beta actin - by Bioz Stars, 2020-01
    90/100 stars

    Images

    1) Product Images from "Support for the initial attachment, growth and differentiation of MG-63 cells: a comparison between nano-size hydroxyapatite and micro-size hydroxyapatite in composites"

    Article Title: Support for the initial attachment, growth and differentiation of MG-63 cells: a comparison between nano-size hydroxyapatite and micro-size hydroxyapatite in composites

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S56661

    Concentrations (percentage of PS) of beta-actin ( A ), vinculin ( B ), talin ( C ). osteocalcin ( D ), osteopontin ( E ), and ICAM ( F ) in MG-63 cells growing on composite material (0), composite materials with 2, 5,10, 15, and 25 vol% of nano-sized tricalcium phosphate (N2–N25) or of micro-sized tricalcium phosphate (M2–M25) or on control cell culture PS on day 3 after seeding. Note: Data presented as mean ± standard error of mean, from 5–9 measurements. Abbreviations: ICAM, intercellular adhesion molecule; PS, polystyrene.
    Figure Legend Snippet: Concentrations (percentage of PS) of beta-actin ( A ), vinculin ( B ), talin ( C ). osteocalcin ( D ), osteopontin ( E ), and ICAM ( F ) in MG-63 cells growing on composite material (0), composite materials with 2, 5,10, 15, and 25 vol% of nano-sized tricalcium phosphate (N2–N25) or of micro-sized tricalcium phosphate (M2–M25) or on control cell culture PS on day 3 after seeding. Note: Data presented as mean ± standard error of mean, from 5–9 measurements. Abbreviations: ICAM, intercellular adhesion molecule; PS, polystyrene.

    Techniques Used: Cell Culture

    Related Articles

    Centrifugation:

    Article Title: Modulating BAP1 expression affects ROS homeostasis, cell motility and mitochondrial function
    Article Snippet: For cytoplasmic and nuclear protein fractions, cells were collected by centrifugation at 1,500 rpm for 5 minutes and washed in phosphate buffered saline (PBS). .. After blocking with blocking buffer (LI-COR, Lincoln, NE, USA), mixed with 0.1% Tween-20, the membrane was incubated with primary antibodies overnight at 4°C (BAP1, Santa Cruz sc-4 sc-28383, 1/500; CDH2, Cell signaling #14215, 1/200; β-actin, Sigma-Aldrich A-5316, 1/10000, PGC1-α, Santa Cruz sc-13067, 1/500, BAF47, BD Pharmingen #612111, 1/1000, OXR1, Novus Biologicals NBP1-86393, 1/1000).

    Positive Control:

    Article Title: Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study
    Article Snippet: Western Blotting At the endpoint of the culture, both cell/scaffolds constructs (n = 3) and controls (scaffolds without cells as a negative control, and cells grown in monolayer up to 80% confluence—4 days—as a positive control) were washed in D-PBS and resuspended in a cell lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 2 mM EDTA, 1% NP-40, and antiproteases 1×, all from Sigma Aldrich). .. The membrane was blocked with 4% dry non-fat milk in 0.1% Tween/Tris Buffer Saline (T-TBS) and incubated overnight at 4 °C with anti β-actin mouse monoclonal antibody (A-5316, Sigma-Aldrich) or anti-HPRT rabbit monoclonal antibody (ab109021, Abcam, Cambridge, UK) diluted 1:2000 or 1:10,000 in T-TBS, respectively.

    Construct:

    Article Title: Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study
    Article Snippet: Western Blotting At the endpoint of the culture, both cell/scaffolds constructs (n = 3) and controls (scaffolds without cells as a negative control, and cells grown in monolayer up to 80% confluence—4 days—as a positive control) were washed in D-PBS and resuspended in a cell lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 2 mM EDTA, 1% NP-40, and antiproteases 1×, all from Sigma Aldrich). .. The membrane was blocked with 4% dry non-fat milk in 0.1% Tween/Tris Buffer Saline (T-TBS) and incubated overnight at 4 °C with anti β-actin mouse monoclonal antibody (A-5316, Sigma-Aldrich) or anti-HPRT rabbit monoclonal antibody (ab109021, Abcam, Cambridge, UK) diluted 1:2000 or 1:10,000 in T-TBS, respectively.

    Electrophoresis:

    Article Title: Transcriptomic analysis reveals sex-specific differences in the expression of Dcl1 and Fis1 genes in the radio-adaptive response of thymocytes to TRP53-mediated apoptosis
    Article Snippet: Electrophoresis was performed using 4–15 % Mini-PROTEAN TGX™ precast gels (BioRad, Hercules, CA). .. For β-actin (A 5316; Sigma) was used a dilution of 1:5000.

    Article Title: Effects of Balance and Gait Training on the Recovery of the Motor Function in an Animal Model of Parkinson's Disease
    Article Snippet: The extracted protein was loaded onto the same amount of SDS polyacrylamide gel and for electrophoresis. .. To confirm if the equivalent protein, monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA), had been loaded, goat anti-mouse IgG (BD Bioscience, Flanklin Lakes, NJ, USA) was used for comparison.

    Article Title: High fat diet increases melanoma cell growth in the bone marrow by inducing osteopontin and interleukin 6
    Article Snippet: Western blot analysis Electrophoresis of 15 mg protein was performed. .. Following protein transfer into 0.45 μM nitrocellulose (NC) membrane (BD, Cat.#1620115), membranes were blocked with 5% skim-milk in TBS-T buffer (0.05%) for 2 h. Then, membranes that washed and were probed using the following antibodies: Nuclear factor kappa B (NF-κB) p65 (Cell signaling Technology, Inc. Cat.#8242; 1:500 dilutions), phospho-NF-κB p65 (Ser468) (Cell signaling Technology, Inc. Cat.#3039; 1:500 dilutions) and anti–β-actin mouse mAb (Sigma-Aldrich, Inc. Cat.#A5316; 1:5,000 dilutions), and the secondary antibodies.

    Article Title: Beneficial Effects of Antecedent Exercise Training on Limb Motor Function and Calpain Expression in a Rat Model of Stroke
    Article Snippet: After electrophoresis, the proteins were transferred onto polyvinylidene fluoride membranes and nonspecific binding was blocked with 5% nonfat dry milk in tris-buffered saline and Tween 20. .. After incubation with the rabbit polyclonal anti-m-calpain antibody (1:1,000 Triple Point Biologics; Forest Grove, OR, USA) and the monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibody, the membranes were washed with TBST and incubated with appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution).

    Article Title: The Effects of Antecedent Exercise on Motor Function Recovery and Brain-derived Neurotrophic Factor Expression after Focal Cerebral Ischemia in Rats
    Article Snippet: After electrophoresis, the proteins were transferred onto polyvinylidene fluoride (PVDF) membranes and nonspecific binding was blocked with 5% nonfat dry milk in tris-buffered saline and Tween 20. .. After incubation with the anti-BDNF primary antibody (sc-546, Santa Cruz, USA), monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibodies membranes were washed with TBST and incubated with the appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution).

    Article Title: Glutamate-induced and NMDA receptor-mediated neurodegeneration entails P2Y1 receptor activation
    Article Snippet: Protein extracts from cultured hippocampal neurons or hippocampal homogenates were diluted in 6× sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer ((0.5 M Tris, 0.4% SDS, pH 6.8), 30% glycerol, 10% SDS, 0.6 M dithiothreitol, and 0.012% of bromophenol blue), boiled at 95 °C for 5 min and then separated by 10% SDS-PAGE electrophoresis. .. A rabbit antibody directed against GAPDH (glyceraldehyde 3-phosphate dehydrogenase) (1:1000; Abcam, Cat#ab9485 RRID:AB_307275) and mouse monoclonal antibodies against α-tubulin (1:20 000; Sigma, Cat#T6074 RRID:AB_477582) or β-actin (1:20,000; Sigma, Cat#A5316 RRID:AB_476743) were used as loading controls.

    Incubation:

    Article Title: Modulating BAP1 expression affects ROS homeostasis, cell motility and mitochondrial function
    Article Snippet: .. After blocking with blocking buffer (LI-COR, Lincoln, NE, USA), mixed with 0.1% Tween-20, the membrane was incubated with primary antibodies overnight at 4°C (BAP1, Santa Cruz sc-4 sc-28383, 1/500; CDH2, Cell signaling #14215, 1/200; β-actin, Sigma-Aldrich A-5316, 1/10000, PGC1-α, Santa Cruz sc-13067, 1/500, BAF47, BD Pharmingen #612111, 1/1000, OXR1, Novus Biologicals NBP1-86393, 1/1000). .. Following incubation with appropriate fluorescent secondary antibody (1/10 000 mixed with 0.1% Tween-20), the immunoreactive bands were visualized using LI-COR-Odyssey infra-red scanner (LI-COR).

    Article Title: Transcriptomic analysis reveals sex-specific differences in the expression of Dcl1 and Fis1 genes in the radio-adaptive response of thymocytes to TRP53-mediated apoptosis
    Article Snippet: After blocking, membranes were incubated with each primary antibody in either 5 % w/v BSA or nonfat dry milk, 1× TBS, 0.1 % Tween 20 overnight at 4 °C. .. For β-actin (A 5316; Sigma) was used a dilution of 1:5000.

    Article Title: Beneficial Effects of Antecedent Exercise Training on Limb Motor Function and Calpain Expression in a Rat Model of Stroke
    Article Snippet: .. After incubation with the rabbit polyclonal anti-m-calpain antibody (1:1,000 Triple Point Biologics; Forest Grove, OR, USA) and the monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibody, the membranes were washed with TBST and incubated with appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution). ..

    Article Title: Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study
    Article Snippet: .. The membrane was blocked with 4% dry non-fat milk in 0.1% Tween/Tris Buffer Saline (T-TBS) and incubated overnight at 4 °C with anti β-actin mouse monoclonal antibody (A-5316, Sigma-Aldrich) or anti-HPRT rabbit monoclonal antibody (ab109021, Abcam, Cambridge, UK) diluted 1:2000 or 1:10,000 in T-TBS, respectively. .. Anti-mouse or anti-rabbit Horse Radish Peroxidase (HRP)-conjugated antibodies (KPL, Gaithersburg, MD, USA), both diluted 1:2000 in 4% dry non-fat milk in T-TBS for 1 h at room temperature, were used as secondary antibodies and the immunocomplexes were detected by chemiluminescence (ECL clarity, BioRad) using Chemi-Doc XRS+ (BioRad).

    Article Title: The Effects of Antecedent Exercise on Motor Function Recovery and Brain-derived Neurotrophic Factor Expression after Focal Cerebral Ischemia in Rats
    Article Snippet: .. After incubation with the anti-BDNF primary antibody (sc-546, Santa Cruz, USA), monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibodies membranes were washed with TBST and incubated with the appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution). ..

    Article Title: Normal expression of DNA repair proteins, hMre11, Rad50 and Rad51 but protracted formation of Rad50 containing foci in X-irradiated skin fibroblasts from radiosensitive cancer patients
    Article Snippet: Equal loading and transfer were assessed by reprobing the blots with anti-β -actin antibody (Sigma A-5316) and Ponceau red (Sigma 19 976-1), respectively. β -Actin was used as an internal standard to account for variations in the amount of protein (usually 20 μ g) loaded in each lane. .. For hMre11, Rad51 or Rad50 detection, membranes were incubated with respective primary and species-specific peroxidase-labelled secondary antibodies according to standard procedures.

    Article Title: Glutamate-induced and NMDA receptor-mediated neurodegeneration entails P2Y1 receptor activation
    Article Snippet: A rabbit antibody directed against GAPDH (glyceraldehyde 3-phosphate dehydrogenase) (1:1000; Abcam, Cat#ab9485 RRID:AB_307275) and mouse monoclonal antibodies against α-tubulin (1:20 000; Sigma, Cat#T6074 RRID:AB_477582) or β-actin (1:20,000; Sigma, Cat#A5316 RRID:AB_476743) were used as loading controls. .. The membranes were incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit or goat anti-mouse or goat anti-guinea-pig secondary antibodies (1:10,000; Pierce) and subsequently with SuperSignal West Pico Chemiluminescent Substrate (Pierce, Alfagene) or with Luminata Forte Western HRP Substrate (Millipore).

    Expressing:

    Article Title: Effects of Balance and Gait Training on the Recovery of the Motor Function in an Animal Model of Parkinson's Disease
    Article Snippet: To confirm if the equivalent protein, monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA), had been loaded, goat anti-mouse IgG (BD Bioscience, Flanklin Lakes, NJ, USA) was used for comparison. .. The significance of differences between the groups’ neurobehavioral scores and the change in TH Protein expression were analyzed using one-way ANOVA, and Tukey’s multiple-range test was used as a post-hoc test.

    Article Title: Normal expression of DNA repair proteins, hMre11, Rad50 and Rad51 but protracted formation of Rad50 containing foci in X-irradiated skin fibroblasts from radiosensitive cancer patients
    Article Snippet: Equal loading and transfer were assessed by reprobing the blots with anti-β -actin antibody (Sigma A-5316) and Ponceau red (Sigma 19 976-1), respectively. β -Actin was used as an internal standard to account for variations in the amount of protein (usually 20 μ g) loaded in each lane. .. The levels of protein expression were quantified using the Kodak 1D Image analysing software (Scientific Imaging Systems, Eastman Kodak Company, Rochester, NY, USA) and normalised to the β -actin levels.

    BIA-KA:

    Article Title: Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study
    Article Snippet: Sample lysates were centrifuged at 15,000 rpm for 20 min at 4 °C and protein concentration in the supernatants was determined by the bicinchoninic acid (BCA; Pierce, Rockford, IL, USA) microplate method. .. The membrane was blocked with 4% dry non-fat milk in 0.1% Tween/Tris Buffer Saline (T-TBS) and incubated overnight at 4 °C with anti β-actin mouse monoclonal antibody (A-5316, Sigma-Aldrich) or anti-HPRT rabbit monoclonal antibody (ab109021, Abcam, Cambridge, UK) diluted 1:2000 or 1:10,000 in T-TBS, respectively.

    Western Blot:

    Article Title: Lysine methyltransferase G9a is not required for DNMT3A/3B anchoring to methylated nucleosomes and maintenance of DNA methylation in somatic cells
    Article Snippet: Paragraph title: Western blot analysis ... Antibodies against H3 (ab1791), DNMT3A (ab2850), and SUV39h1 (ab12405) were purchased from Abcam Inc. (Cambridge, UK); DNMT1 (sc-20701) and DNMT3B (sc-10235) from Santa Cruz Biotech (Santa Cruz, CA); Myc epitope tag (05-724) from Upstate (now Millipore, Billerica, MA) and G9a (G 6919); β-Actin (A 5316) from Sigma (Saint Louis, MO).

    Article Title: Transcriptomic analysis reveals sex-specific differences in the expression of Dcl1 and Fis1 genes in the radio-adaptive response of thymocytes to TRP53-mediated apoptosis
    Article Snippet: Paragraph title: Western blot analysis ... For β-actin (A 5316; Sigma) was used a dilution of 1:5000.

    Article Title: LXCXE-independent chromatin remodeling by Rb/E2f mediates neuronal quiescence
    Article Snippet: .. Protein was isolated from cultured cortical neurons and western blot analyses performed as previously described, with antibodies directed toward cleaved caspase-3 (Cell Signaling, Cat No. 9664S), Rb (PharMingen, Cat No. 554136), Cyclin A2 (Abcam, Cat No. ab7965), PCNA (Millpore, Cat No. MAB424) and β-actin (Sigma, Cat No. A-5316). ..

    Article Title: High fat diet increases melanoma cell growth in the bone marrow by inducing osteopontin and interleukin 6
    Article Snippet: Paragraph title: Western blot analysis ... Following protein transfer into 0.45 μM nitrocellulose (NC) membrane (BD, Cat.#1620115), membranes were blocked with 5% skim-milk in TBS-T buffer (0.05%) for 2 h. Then, membranes that washed and were probed using the following antibodies: Nuclear factor kappa B (NF-κB) p65 (Cell signaling Technology, Inc. Cat.#8242; 1:500 dilutions), phospho-NF-κB p65 (Ser468) (Cell signaling Technology, Inc. Cat.#3039; 1:500 dilutions) and anti–β-actin mouse mAb (Sigma-Aldrich, Inc. Cat.#A5316; 1:5,000 dilutions), and the secondary antibodies.

    Article Title: Beneficial Effects of Antecedent Exercise Training on Limb Motor Function and Calpain Expression in a Rat Model of Stroke
    Article Snippet: The hippocampus was homogenized in lysis buffer B (137 mM NaCl, 20 mM Tris, 1% NP40, 10% glycerol, 1 mM phenylmethylsulfonyl fluoride, 10 µg/ml aprotinin, 1 µg/ml leupeptin, 0.5 mM sodium vanadate, pH 8.0) for western blot analysis. .. After incubation with the rabbit polyclonal anti-m-calpain antibody (1:1,000 Triple Point Biologics; Forest Grove, OR, USA) and the monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibody, the membranes were washed with TBST and incubated with appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution).

    Article Title: Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study
    Article Snippet: Paragraph title: 4.10. Western Blotting ... The membrane was blocked with 4% dry non-fat milk in 0.1% Tween/Tris Buffer Saline (T-TBS) and incubated overnight at 4 °C with anti β-actin mouse monoclonal antibody (A-5316, Sigma-Aldrich) or anti-HPRT rabbit monoclonal antibody (ab109021, Abcam, Cambridge, UK) diluted 1:2000 or 1:10,000 in T-TBS, respectively.

    Article Title: The Effects of Antecedent Exercise on Motor Function Recovery and Brain-derived Neurotrophic Factor Expression after Focal Cerebral Ischemia in Rats
    Article Snippet: The hippocampus was homogenized in lysis buffer B (137 mM NaCl, 20 mM Tris, 1% NP40, 10% glycerol, 1 mM phenylmethylsulfonyl fluoride [PMSF], 10 µg/Ml aprotinin, 1 µg/Ml leupeptin, 0.5 mM sodium vanadate, pH8.0) for western blot analysis. .. After incubation with the anti-BDNF primary antibody (sc-546, Santa Cruz, USA), monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibodies membranes were washed with TBST and incubated with the appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution).

    Article Title: Escherichia coli Subtilase Cytotoxin Induces Apoptosis Regulated by Host Bcl-2 Family Proteins Bax/Bak ▿
    Article Snippet: Paragraph title: Western immunoblotting. ... Anti-Puma (catalog no. P-4618) and anti-β-actin (AC-74) (catalog no. A-5316) were from Sigma.

    Article Title: Normal expression of DNA repair proteins, hMre11, Rad50 and Rad51 but protracted formation of Rad50 containing foci in X-irradiated skin fibroblasts from radiosensitive cancer patients
    Article Snippet: Paragraph title: Western blot analysis ... Equal loading and transfer were assessed by reprobing the blots with anti-β -actin antibody (Sigma A-5316) and Ponceau red (Sigma 19 976-1), respectively. β -Actin was used as an internal standard to account for variations in the amount of protein (usually 20 μ g) loaded in each lane.

    Article Title: Glutamate-induced and NMDA receptor-mediated neurodegeneration entails P2Y1 receptor activation
    Article Snippet: Paragraph title: Western blot analysis ... A rabbit antibody directed against GAPDH (glyceraldehyde 3-phosphate dehydrogenase) (1:1000; Abcam, Cat#ab9485 RRID:AB_307275) and mouse monoclonal antibodies against α-tubulin (1:20 000; Sigma, Cat#T6074 RRID:AB_477582) or β-actin (1:20,000; Sigma, Cat#A5316 RRID:AB_476743) were used as loading controls.

    Electrotransfer:

    Article Title: Glutamate-induced and NMDA receptor-mediated neurodegeneration entails P2Y1 receptor activation
    Article Snippet: After the electrotransfer of the proteins, CRMP2 protein was detected using a rabbit antibody directed against CRMP2 (1:500; Abcam, Cat#ab36201, RRID:AB_731750) and the different synaptic markers were detected using mouse monoclonal antibodies directed against PSD-95 (1:20,000; Millipore, Merck, Cat#P246, RRID:AB_260911), synaptophysin (1:2000–1:20,000; Sigma, Cat#S5768, RRID:AB_477523), and syntaxin 1 A (1:2000; Sigma, Cat#S0664, RRID:AB_477483), guinea-pig polyclonal anti-vGluT1 (1:10,000; Synaptic Systems, Goettingen, Germany, Cat#135304 RRID:AB_887878), anti-vGAT (1:1000; Synaptic Systems; Cat#131004, RRID:AB_887873), and rabbit polyclonal anti-gephyrin (1:10,000; Abcam, Cat#ab25784 RRID:AB_1209349), all diluted in TBS-T (Tris-buffered saline, 0.1% Tween 20) containing 5% dried milk. .. A rabbit antibody directed against GAPDH (glyceraldehyde 3-phosphate dehydrogenase) (1:1000; Abcam, Cat#ab9485 RRID:AB_307275) and mouse monoclonal antibodies against α-tubulin (1:20 000; Sigma, Cat#T6074 RRID:AB_477582) or β-actin (1:20,000; Sigma, Cat#A5316 RRID:AB_476743) were used as loading controls.

    TCA Precipitation:

    Article Title: Lysine methyltransferase G9a is not required for DNMT3A/3B anchoring to methylated nucleosomes and maintenance of DNA methylation in somatic cells
    Article Snippet: Western blot analysis Proteins from the same volume of each fraction (200 to 250 μl) were concentrated by TCA precipitation, dissolved in SDS/β-mercaptoethanol loading buffer, and resolved on a 4% to 15% gradient SDS/PAGE gel (Bio-Rad, Hercules, CA). .. Antibodies against H3 (ab1791), DNMT3A (ab2850), and SUV39h1 (ab12405) were purchased from Abcam Inc. (Cambridge, UK); DNMT1 (sc-20701) and DNMT3B (sc-10235) from Santa Cruz Biotech (Santa Cruz, CA); Myc epitope tag (05-724) from Upstate (now Millipore, Billerica, MA) and G9a (G 6919); β-Actin (A 5316) from Sigma (Saint Louis, MO).

    Cell Culture:

    Article Title: LXCXE-independent chromatin remodeling by Rb/E2f mediates neuronal quiescence
    Article Snippet: .. Protein was isolated from cultured cortical neurons and western blot analyses performed as previously described, with antibodies directed toward cleaved caspase-3 (Cell Signaling, Cat No. 9664S), Rb (PharMingen, Cat No. 554136), Cyclin A2 (Abcam, Cat No. ab7965), PCNA (Millpore, Cat No. MAB424) and β-actin (Sigma, Cat No. A-5316). ..

    Article Title: Glutamate-induced and NMDA receptor-mediated neurodegeneration entails P2Y1 receptor activation
    Article Snippet: Protein extracts from cultured hippocampal neurons or hippocampal homogenates were diluted in 6× sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer ((0.5 M Tris, 0.4% SDS, pH 6.8), 30% glycerol, 10% SDS, 0.6 M dithiothreitol, and 0.012% of bromophenol blue), boiled at 95 °C for 5 min and then separated by 10% SDS-PAGE electrophoresis. .. A rabbit antibody directed against GAPDH (glyceraldehyde 3-phosphate dehydrogenase) (1:1000; Abcam, Cat#ab9485 RRID:AB_307275) and mouse monoclonal antibodies against α-tubulin (1:20 000; Sigma, Cat#T6074 RRID:AB_477582) or β-actin (1:20,000; Sigma, Cat#A5316 RRID:AB_476743) were used as loading controls.

    other:

    Article Title: Cleavage of DFNA5 by caspase-3 during apoptosis mediates progression to secondary necrotic/pyroptotic cell death
    Article Snippet: Monoclonal anti-β-actin (Catalogue No. A-5316) was from Sigma Aldrich.

    Imaging:

    Article Title: Escherichia coli Subtilase Cytotoxin Induces Apoptosis Regulated by Host Bcl-2 Family Proteins Bax/Bak ▿
    Article Snippet: Anti-Puma (catalog no. P-4618) and anti-β-actin (AC-74) (catalog no. A-5316) were from Sigma. .. Images were captured using a Kodak 4000MM image station (Carestream Molecular Imaging), and net band intensities (mean pixel intensity by number of pixels) were determined using MI imaging software (Carestream Molecular Imaging).

    Article Title: Normal expression of DNA repair proteins, hMre11, Rad50 and Rad51 but protracted formation of Rad50 containing foci in X-irradiated skin fibroblasts from radiosensitive cancer patients
    Article Snippet: Equal loading and transfer were assessed by reprobing the blots with anti-β -actin antibody (Sigma A-5316) and Ponceau red (Sigma 19 976-1), respectively. β -Actin was used as an internal standard to account for variations in the amount of protein (usually 20 μ g) loaded in each lane. .. The levels of protein expression were quantified using the Kodak 1D Image analysing software (Scientific Imaging Systems, Eastman Kodak Company, Rochester, NY, USA) and normalised to the β -actin levels.

    Protein Concentration:

    Article Title: Effects of Balance and Gait Training on the Recovery of the Motor Function in an Animal Model of Parkinson's Disease
    Article Snippet: The samples were centrifuged for 20 min at 15,000 rpm to obtain supernatant, and the protein concentration in the solution was quantified using Bradford method (Bio-Rad protein assay). .. To confirm if the equivalent protein, monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA), had been loaded, goat anti-mouse IgG (BD Bioscience, Flanklin Lakes, NJ, USA) was used for comparison.

    Article Title: Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study
    Article Snippet: Sample lysates were centrifuged at 15,000 rpm for 20 min at 4 °C and protein concentration in the supernatants was determined by the bicinchoninic acid (BCA; Pierce, Rockford, IL, USA) microplate method. .. The membrane was blocked with 4% dry non-fat milk in 0.1% Tween/Tris Buffer Saline (T-TBS) and incubated overnight at 4 °C with anti β-actin mouse monoclonal antibody (A-5316, Sigma-Aldrich) or anti-HPRT rabbit monoclonal antibody (ab109021, Abcam, Cambridge, UK) diluted 1:2000 or 1:10,000 in T-TBS, respectively.

    Binding Assay:

    Article Title: Effects of Balance and Gait Training on the Recovery of the Motor Function in an Animal Model of Parkinson's Disease
    Article Snippet: Thereafter, the gel was transferred to a nitrocellulose membrane (Whatman GmbH, Dassel, Germany) and underwent reacted with TBX and 5% non fat dried milk for 1 hr at 4°C to prevent non-specific binding with the antibody. .. To confirm if the equivalent protein, monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA), had been loaded, goat anti-mouse IgG (BD Bioscience, Flanklin Lakes, NJ, USA) was used for comparison.

    Article Title: Beneficial Effects of Antecedent Exercise Training on Limb Motor Function and Calpain Expression in a Rat Model of Stroke
    Article Snippet: After electrophoresis, the proteins were transferred onto polyvinylidene fluoride membranes and nonspecific binding was blocked with 5% nonfat dry milk in tris-buffered saline and Tween 20. .. After incubation with the rabbit polyclonal anti-m-calpain antibody (1:1,000 Triple Point Biologics; Forest Grove, OR, USA) and the monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibody, the membranes were washed with TBST and incubated with appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution).

    Nucleic Acid Electrophoresis:

    Article Title: Glutamate-induced and NMDA receptor-mediated neurodegeneration entails P2Y1 receptor activation
    Article Snippet: Protein extracts from cultured hippocampal neurons or hippocampal homogenates were diluted in 6× sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer ((0.5 M Tris, 0.4% SDS, pH 6.8), 30% glycerol, 10% SDS, 0.6 M dithiothreitol, and 0.012% of bromophenol blue), boiled at 95 °C for 5 min and then separated by 10% SDS-PAGE electrophoresis. .. A rabbit antibody directed against GAPDH (glyceraldehyde 3-phosphate dehydrogenase) (1:1000; Abcam, Cat#ab9485 RRID:AB_307275) and mouse monoclonal antibodies against α-tubulin (1:20 000; Sigma, Cat#T6074 RRID:AB_477582) or β-actin (1:20,000; Sigma, Cat#A5316 RRID:AB_476743) were used as loading controls.

    Isolation:

    Article Title: LXCXE-independent chromatin remodeling by Rb/E2f mediates neuronal quiescence
    Article Snippet: .. Protein was isolated from cultured cortical neurons and western blot analyses performed as previously described, with antibodies directed toward cleaved caspase-3 (Cell Signaling, Cat No. 9664S), Rb (PharMingen, Cat No. 554136), Cyclin A2 (Abcam, Cat No. ab7965), PCNA (Millpore, Cat No. MAB424) and β-actin (Sigma, Cat No. A-5316). ..

    Protein Extraction:

    Article Title: Modulating BAP1 expression affects ROS homeostasis, cell motility and mitochondrial function
    Article Snippet: Paragraph title: Protein extraction and quantification ... After blocking with blocking buffer (LI-COR, Lincoln, NE, USA), mixed with 0.1% Tween-20, the membrane was incubated with primary antibodies overnight at 4°C (BAP1, Santa Cruz sc-4 sc-28383, 1/500; CDH2, Cell signaling #14215, 1/200; β-actin, Sigma-Aldrich A-5316, 1/10000, PGC1-α, Santa Cruz sc-13067, 1/500, BAF47, BD Pharmingen #612111, 1/1000, OXR1, Novus Biologicals NBP1-86393, 1/1000).

    Blocking Assay:

    Article Title: Modulating BAP1 expression affects ROS homeostasis, cell motility and mitochondrial function
    Article Snippet: .. After blocking with blocking buffer (LI-COR, Lincoln, NE, USA), mixed with 0.1% Tween-20, the membrane was incubated with primary antibodies overnight at 4°C (BAP1, Santa Cruz sc-4 sc-28383, 1/500; CDH2, Cell signaling #14215, 1/200; β-actin, Sigma-Aldrich A-5316, 1/10000, PGC1-α, Santa Cruz sc-13067, 1/500, BAF47, BD Pharmingen #612111, 1/1000, OXR1, Novus Biologicals NBP1-86393, 1/1000). .. Following incubation with appropriate fluorescent secondary antibody (1/10 000 mixed with 0.1% Tween-20), the immunoreactive bands were visualized using LI-COR-Odyssey infra-red scanner (LI-COR).

    Article Title: Transcriptomic analysis reveals sex-specific differences in the expression of Dcl1 and Fis1 genes in the radio-adaptive response of thymocytes to TRP53-mediated apoptosis
    Article Snippet: After blocking, membranes were incubated with each primary antibody in either 5 % w/v BSA or nonfat dry milk, 1× TBS, 0.1 % Tween 20 overnight at 4 °C. .. For β-actin (A 5316; Sigma) was used a dilution of 1:5000.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: High fat diet increases melanoma cell growth in the bone marrow by inducing osteopontin and interleukin 6
    Article Snippet: .. Following protein transfer into 0.45 μM nitrocellulose (NC) membrane (BD, Cat.#1620115), membranes were blocked with 5% skim-milk in TBS-T buffer (0.05%) for 2 h. Then, membranes that washed and were probed using the following antibodies: Nuclear factor kappa B (NF-κB) p65 (Cell signaling Technology, Inc. Cat.#8242; 1:500 dilutions), phospho-NF-κB p65 (Ser468) (Cell signaling Technology, Inc. Cat.#3039; 1:500 dilutions) and anti–β-actin mouse mAb (Sigma-Aldrich, Inc. Cat.#A5316; 1:5,000 dilutions), and the secondary antibodies. .. Chemiluminescent reagent (GE Healthcare, Cat.#RPN2106) and detected on Amersham Hyperfilm ECL (GE Healthcare, Cat.#28906839) were used for the revelation of the western blot.

    Article Title: Oncogenic activation of the RNA binding protein NELFE and MYC signaling in hepatocellular carcinoma
    Article Snippet: .. Protein detection was performed using anti-NELFE (Abcam, cat# ab170104), anti-β–actin (Sigma-Aldrich, catA5316), anti-Cyclin B1 (Cell Signaling Technology, cat #4138), anti-HLA-A (Abcam, cat#52922), anti-HRAS (Santa Cruz, cat#sc-520) and anti-SV40 T Ag (Santa Cruz, cat#sc-147). ..

    Article Title: Glutamate-induced and NMDA receptor-mediated neurodegeneration entails P2Y1 receptor activation
    Article Snippet: .. A rabbit antibody directed against GAPDH (glyceraldehyde 3-phosphate dehydrogenase) (1:1000; Abcam, Catab9485 RRID:AB_307275) and mouse monoclonal antibodies against α-tubulin (1:20 000; Sigma, Cat#T6074 RRID:AB_477582) or β-actin (1:20,000; Sigma, Cat#A5316 RRID:AB_476743) were used as loading controls. .. The membranes were incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit or goat anti-mouse or goat anti-guinea-pig secondary antibodies (1:10,000; Pierce) and subsequently with SuperSignal West Pico Chemiluminescent Substrate (Pierce, Alfagene) or with Luminata Forte Western HRP Substrate (Millipore).

    SDS Page:

    Article Title: Lysine methyltransferase G9a is not required for DNMT3A/3B anchoring to methylated nucleosomes and maintenance of DNA methylation in somatic cells
    Article Snippet: Western blot analysis Proteins from the same volume of each fraction (200 to 250 μl) were concentrated by TCA precipitation, dissolved in SDS/β-mercaptoethanol loading buffer, and resolved on a 4% to 15% gradient SDS/PAGE gel (Bio-Rad, Hercules, CA). .. Antibodies against H3 (ab1791), DNMT3A (ab2850), and SUV39h1 (ab12405) were purchased from Abcam Inc. (Cambridge, UK); DNMT1 (sc-20701) and DNMT3B (sc-10235) from Santa Cruz Biotech (Santa Cruz, CA); Myc epitope tag (05-724) from Upstate (now Millipore, Billerica, MA) and G9a (G 6919); β-Actin (A 5316) from Sigma (Saint Louis, MO).

    Article Title: Modulating BAP1 expression affects ROS homeostasis, cell motility and mitochondrial function
    Article Snippet: 50μg of protein extracts were resolved by 4-15% SDS-PAGE (SDS-PAGE Mini- PROTEAN® TGX™ #456-1086 Biorad) and transferred to 0.2 μm nitrocellulose membranes (Biorad). .. After blocking with blocking buffer (LI-COR, Lincoln, NE, USA), mixed with 0.1% Tween-20, the membrane was incubated with primary antibodies overnight at 4°C (BAP1, Santa Cruz sc-4 sc-28383, 1/500; CDH2, Cell signaling #14215, 1/200; β-actin, Sigma-Aldrich A-5316, 1/10000, PGC1-α, Santa Cruz sc-13067, 1/500, BAF47, BD Pharmingen #612111, 1/1000, OXR1, Novus Biologicals NBP1-86393, 1/1000).

    Article Title: Glutamate-induced and NMDA receptor-mediated neurodegeneration entails P2Y1 receptor activation
    Article Snippet: Protein extracts from cultured hippocampal neurons or hippocampal homogenates were diluted in 6× sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer ((0.5 M Tris, 0.4% SDS, pH 6.8), 30% glycerol, 10% SDS, 0.6 M dithiothreitol, and 0.012% of bromophenol blue), boiled at 95 °C for 5 min and then separated by 10% SDS-PAGE electrophoresis. .. A rabbit antibody directed against GAPDH (glyceraldehyde 3-phosphate dehydrogenase) (1:1000; Abcam, Cat#ab9485 RRID:AB_307275) and mouse monoclonal antibodies against α-tubulin (1:20 000; Sigma, Cat#T6074 RRID:AB_477582) or β-actin (1:20,000; Sigma, Cat#A5316 RRID:AB_476743) were used as loading controls.

    Plasmid Preparation:

    Article Title: A polymorphism in the lysyl oxidase propeptide domain accelerates carcinogen-induced cancer
    Article Snippet: .. Antibodies used were Ki-67 (cloneSP6; Thermo Scientific), LOX-PP (rabbit polyclonal IgG) used as described previously , β-actin (#A-5316; Sigma–Aldrich) and biotinylated goat anti-rabbit IgG (Vector). ..

    Software:

    Article Title: Beneficial Effects of Antecedent Exercise Training on Limb Motor Function and Calpain Expression in a Rat Model of Stroke
    Article Snippet: After incubation with the rabbit polyclonal anti-m-calpain antibody (1:1,000 Triple Point Biologics; Forest Grove, OR, USA) and the monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibody, the membranes were washed with TBST and incubated with appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution). .. The film signals were digitally scanned and then quantified with NIH image J software.

    Article Title: The Effects of Antecedent Exercise on Motor Function Recovery and Brain-derived Neurotrophic Factor Expression after Focal Cerebral Ischemia in Rats
    Article Snippet: After incubation with the anti-BDNF primary antibody (sc-546, Santa Cruz, USA), monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibodies membranes were washed with TBST and incubated with the appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution). .. The film signals were digitally scanned and then quantified using NIH image J software.

    Article Title: Escherichia coli Subtilase Cytotoxin Induces Apoptosis Regulated by Host Bcl-2 Family Proteins Bax/Bak ▿
    Article Snippet: Anti-Puma (catalog no. P-4618) and anti-β-actin (AC-74) (catalog no. A-5316) were from Sigma. .. Images were captured using a Kodak 4000MM image station (Carestream Molecular Imaging), and net band intensities (mean pixel intensity by number of pixels) were determined using MI imaging software (Carestream Molecular Imaging).

    Article Title: Normal expression of DNA repair proteins, hMre11, Rad50 and Rad51 but protracted formation of Rad50 containing foci in X-irradiated skin fibroblasts from radiosensitive cancer patients
    Article Snippet: Equal loading and transfer were assessed by reprobing the blots with anti-β -actin antibody (Sigma A-5316) and Ponceau red (Sigma 19 976-1), respectively. β -Actin was used as an internal standard to account for variations in the amount of protein (usually 20 μ g) loaded in each lane. .. The levels of protein expression were quantified using the Kodak 1D Image analysing software (Scientific Imaging Systems, Eastman Kodak Company, Rochester, NY, USA) and normalised to the β -actin levels.

    Article Title: Glutamate-induced and NMDA receptor-mediated neurodegeneration entails P2Y1 receptor activation
    Article Snippet: A rabbit antibody directed against GAPDH (glyceraldehyde 3-phosphate dehydrogenase) (1:1000; Abcam, Cat#ab9485 RRID:AB_307275) and mouse monoclonal antibodies against α-tubulin (1:20 000; Sigma, Cat#T6074 RRID:AB_477582) or β-actin (1:20,000; Sigma, Cat#A5316 RRID:AB_476743) were used as loading controls. .. The immunoreactivity was visualized using a VersaDoc3000 apparatus and analyzed using the ImageLab software (BioRad, Amadora, Portugal).

    Irradiation:

    Article Title: Normal expression of DNA repair proteins, hMre11, Rad50 and Rad51 but protracted formation of Rad50 containing foci in X-irradiated skin fibroblasts from radiosensitive cancer patients
    Article Snippet: Western blot analysis Nearly confluent nonirradiated and irradiated with 8 Gy fibroblasts were detached by trypsinisation, lyzed in RIPA buffer (10 mM tris-HCl, pH 7.4, 150 mM NaCl, 1% sodium deoxycholate, 1% Triton X-100) containing protease inhibitors (2 μ g ml−1 aprotinin, 2 μ g ml−1 leupeptin, 5 μ g ml−1 pepstatin, 1 mM phenylmethane sulphonyl fluoride) for 30 min on ice and subsequently centrifuged 10 min at 500 g . .. Equal loading and transfer were assessed by reprobing the blots with anti-β -actin antibody (Sigma A-5316) and Ponceau red (Sigma 19 976-1), respectively. β -Actin was used as an internal standard to account for variations in the amount of protein (usually 20 μ g) loaded in each lane.

    Negative Control:

    Article Title: Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study
    Article Snippet: Western Blotting At the endpoint of the culture, both cell/scaffolds constructs (n = 3) and controls (scaffolds without cells as a negative control, and cells grown in monolayer up to 80% confluence—4 days—as a positive control) were washed in D-PBS and resuspended in a cell lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 2 mM EDTA, 1% NP-40, and antiproteases 1×, all from Sigma Aldrich). .. The membrane was blocked with 4% dry non-fat milk in 0.1% Tween/Tris Buffer Saline (T-TBS) and incubated overnight at 4 °C with anti β-actin mouse monoclonal antibody (A-5316, Sigma-Aldrich) or anti-HPRT rabbit monoclonal antibody (ab109021, Abcam, Cambridge, UK) diluted 1:2000 or 1:10,000 in T-TBS, respectively.

    Radio Immunoprecipitation:

    Article Title: Modulating BAP1 expression affects ROS homeostasis, cell motility and mitochondrial function
    Article Snippet: Cell pellets were lysed in radioimmunoprecipitation assay buffer (RIPA) [100mM Tris-HCl/pH7.5, 0.1M NaCl, 1mM EDTA, 1% Triton, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS)], supplemented with NaF, Na3 VO4 , phenylmethanesulfonylfluoride (PMSF) and a cocktail of protease inhibitors (cOmplete©, Roche). .. After blocking with blocking buffer (LI-COR, Lincoln, NE, USA), mixed with 0.1% Tween-20, the membrane was incubated with primary antibodies overnight at 4°C (BAP1, Santa Cruz sc-4 sc-28383, 1/500; CDH2, Cell signaling #14215, 1/200; β-actin, Sigma-Aldrich A-5316, 1/10000, PGC1-α, Santa Cruz sc-13067, 1/500, BAF47, BD Pharmingen #612111, 1/1000, OXR1, Novus Biologicals NBP1-86393, 1/1000).

    Lysis:

    Article Title: Beneficial Effects of Antecedent Exercise Training on Limb Motor Function and Calpain Expression in a Rat Model of Stroke
    Article Snippet: The tissue was homogenized in freshly prepared lysis buffer (1:10 w/v) and centrifuged at 12,000 × g for 30 min. .. After incubation with the rabbit polyclonal anti-m-calpain antibody (1:1,000 Triple Point Biologics; Forest Grove, OR, USA) and the monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibody, the membranes were washed with TBST and incubated with appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution).

    Article Title: Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study
    Article Snippet: Western Blotting At the endpoint of the culture, both cell/scaffolds constructs (n = 3) and controls (scaffolds without cells as a negative control, and cells grown in monolayer up to 80% confluence—4 days—as a positive control) were washed in D-PBS and resuspended in a cell lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 2 mM EDTA, 1% NP-40, and antiproteases 1×, all from Sigma Aldrich). .. The membrane was blocked with 4% dry non-fat milk in 0.1% Tween/Tris Buffer Saline (T-TBS) and incubated overnight at 4 °C with anti β-actin mouse monoclonal antibody (A-5316, Sigma-Aldrich) or anti-HPRT rabbit monoclonal antibody (ab109021, Abcam, Cambridge, UK) diluted 1:2000 or 1:10,000 in T-TBS, respectively.

    Article Title: The Effects of Antecedent Exercise on Motor Function Recovery and Brain-derived Neurotrophic Factor Expression after Focal Cerebral Ischemia in Rats
    Article Snippet: Tissue was homogenized in freshly prepared lysis buffer (1:10 w/v) and centrifuged at 12,000 ×g for 30 min. .. After incubation with the anti-BDNF primary antibody (sc-546, Santa Cruz, USA), monoclonal anti-β actin (A-5316, 1:5,000, Sigma, USA) antibodies membranes were washed with TBST and incubated with the appropriate horse radish peroxidase-conjugated secondary antibody (1:4,000 dilution).

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    Millipore a5228
    A5228, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a5228/product/Millipore
    Average 90 stars, based on 1 article reviews
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    a5228 - by Bioz Stars, 2020-01
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    90
    Millipore β actin
    Deletion of Arhgef1 inhibits leukocyte rolling and adhesion. ( A ) Time-dependent in vivo effect of Ang II (30 pmol) on leukocyte rolling and adhesion in mesenteric vessels of Arhgef1 lox/lox and Arhgef1 –/– mice ( n = 5 mice). ( B ) Effect of losartan on leukocyte rolling and adhesion induced by Ang II (30 pmol, 4 hours) in mesenteric vessels of Arhgef1 lox/lox and Arhgef1 –/– mice ( n = 5 mice). ( C ) Representative immunoblot of VCAM1, ICAM1, and <t>β-actin</t> in lysates of aortas from Arhgef1 lox/lox and Arhgef1 –/– mice before (0) and 4 and 8 hours after Ang II treatment ( n = 3) and corresponding quantification. All lanes were run on the same gel, but lanes 3 and 4 were noncontiguous as indicated by the black dividing line. ( D ) In vitro static adhesion of Arhgef1 lox/lox and Arhgef1 –/– leukocytes on ICAM before (0) and 1 and 4 hours after Ang II treatment ( n = 6 experiments). ( E ) In vitro analysis of Arhgef1 lox/lox and Arhgef1 –/– leukocyte rolling and adhesion on HUVECs under shear flow, before (–) and 4 hours after (+) Ang II treatment ( n = 5). * P
    β Actin, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1329 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β actin/product/Millipore
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    N/A
    The mouse monoclonal antibody HHF35 recognizes muscle specific alpha and gamma actin 42 kDa in various species This antibody stains skeletal smooth and myocardial cells as well as myoepithelial cells
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    Image Search Results


    Deletion of Arhgef1 inhibits leukocyte rolling and adhesion. ( A ) Time-dependent in vivo effect of Ang II (30 pmol) on leukocyte rolling and adhesion in mesenteric vessels of Arhgef1 lox/lox and Arhgef1 –/– mice ( n = 5 mice). ( B ) Effect of losartan on leukocyte rolling and adhesion induced by Ang II (30 pmol, 4 hours) in mesenteric vessels of Arhgef1 lox/lox and Arhgef1 –/– mice ( n = 5 mice). ( C ) Representative immunoblot of VCAM1, ICAM1, and β-actin in lysates of aortas from Arhgef1 lox/lox and Arhgef1 –/– mice before (0) and 4 and 8 hours after Ang II treatment ( n = 3) and corresponding quantification. All lanes were run on the same gel, but lanes 3 and 4 were noncontiguous as indicated by the black dividing line. ( D ) In vitro static adhesion of Arhgef1 lox/lox and Arhgef1 –/– leukocytes on ICAM before (0) and 1 and 4 hours after Ang II treatment ( n = 6 experiments). ( E ) In vitro analysis of Arhgef1 lox/lox and Arhgef1 –/– leukocyte rolling and adhesion on HUVECs under shear flow, before (–) and 4 hours after (+) Ang II treatment ( n = 5). * P

    Journal: The Journal of Clinical Investigation

    Article Title: Leukocyte RhoA exchange factor Arhgef1 mediates vascular inflammation and atherosclerosis

    doi: 10.1172/JCI92702

    Figure Lengend Snippet: Deletion of Arhgef1 inhibits leukocyte rolling and adhesion. ( A ) Time-dependent in vivo effect of Ang II (30 pmol) on leukocyte rolling and adhesion in mesenteric vessels of Arhgef1 lox/lox and Arhgef1 –/– mice ( n = 5 mice). ( B ) Effect of losartan on leukocyte rolling and adhesion induced by Ang II (30 pmol, 4 hours) in mesenteric vessels of Arhgef1 lox/lox and Arhgef1 –/– mice ( n = 5 mice). ( C ) Representative immunoblot of VCAM1, ICAM1, and β-actin in lysates of aortas from Arhgef1 lox/lox and Arhgef1 –/– mice before (0) and 4 and 8 hours after Ang II treatment ( n = 3) and corresponding quantification. All lanes were run on the same gel, but lanes 3 and 4 were noncontiguous as indicated by the black dividing line. ( D ) In vitro static adhesion of Arhgef1 lox/lox and Arhgef1 –/– leukocytes on ICAM before (0) and 1 and 4 hours after Ang II treatment ( n = 6 experiments). ( E ) In vitro analysis of Arhgef1 lox/lox and Arhgef1 –/– leukocyte rolling and adhesion on HUVECs under shear flow, before (–) and 4 hours after (+) Ang II treatment ( n = 5). * P

    Article Snippet: Equal loading was checked by reprobing of the membrane with mAb to β-actin (A-5316, MilliporeSigma).

    Techniques: In Vivo, Mouse Assay, In Vitro, Flow Cytometry