anti glyceraldehyde 3 phosphate dehydrogenase  (Millipore)


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    Structured Review

    Millipore anti glyceraldehyde 3 phosphate dehydrogenase
    Anti Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 88 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti glyceraldehyde 3 phosphate dehydrogenase/product/Millipore
    Average 99 stars, based on 88 article reviews
    Price from $9.99 to $1999.99
    anti glyceraldehyde 3 phosphate dehydrogenase - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Centrifugation:

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: Cells were washed twice in PBS, lysed in 1× Glasgow lysis buffer (GLB; 1% [vol/vol] Triton X-100, 120 mM KCl, 30 mM NaCl, 5 mM MgCl2 , 10% [vol/vol] glycerol, 10 mM PIPES [piperazine-N ,N ′-bis(2-ethanesulfonic acid)]-NaOH, pH 7.2, with protease and phosphatase inhibitors), and clarified by centrifugation at 2,800 × g for 5 min at 4°C, before determining and normalizing the protein concentration by a bicinchoninic acid assay (Pierce). .. The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma).

    Bradford Assay:

    Article Title: Antibodies to cannabinoid type 1 receptor co-react with stomatin-like protein 2 in mouse brain mitochondria
    Article Snippet: The total protein content of all fractions was determined using the Bradford assay. .. The membranes were subsequently immunoblotted with anti-CB1 (guinea pig; Frontier Science, Japan; 1 : 400), anti-SLP-2 (rabbit; 1 : 200; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (mouse; 1 : 700; Chemicon International, Temecula, CA, USA) for load control.

    Blocking Assay:

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: Following separation by SDS-PAGE, proteins were transferred to a polyvinylidene difluoride (PVDF) membrane and blocked in 50% (vol/vol) Odyssey blocking (OB) buffer (LI-COR) in PBS. .. The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma).

    Electrophoresis:

    Article Title: Estradiol Activates PI3K/Akt/GSK3 Pathway Under Chronic Neurodegenerative Conditions Triggered by Perinatal Asphyxia
    Article Snippet: After electrophoresis (120 V for 90 min), proteins were transferred to polyvinylidene difluoride (PVDF) membrane for 2 h at 260 mA as described previously ( ). .. We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Article Title: Antibodies to cannabinoid type 1 receptor co-react with stomatin-like protein 2 in mouse brain mitochondria
    Article Snippet: Based on protein content, 20-μg samples of the cytosolic and mitochondrial fractions were separated using electrophoresis in 4–12% NuPAGE Bis-Tris mini gels (Invitrogen, Carlsbad, CA, USA), and electrophoretically transferred to polyvinylidene fluoride membranes (Bio-Rad Laboratories, Hercules, CA, USA). .. The membranes were subsequently immunoblotted with anti-CB1 (guinea pig; Frontier Science, Japan; 1 : 400), anti-SLP-2 (rabbit; 1 : 200; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (mouse; 1 : 700; Chemicon International, Temecula, CA, USA) for load control.

    Incubation:

    Article Title: Estradiol Activates PI3K/Akt/GSK3 Pathway Under Chronic Neurodegenerative Conditions Triggered by Perinatal Asphyxia
    Article Snippet: Cytoplasmic cellular fractions (n = 4 per group) were incubated overnight at 4°C with the following primary antibodies: anti-GPER (1:1000, mouse-IgG, Sigma-Aldrich), anti-ERα (mouse-IgG, 1:500, Sigma-Aldrich), anti-ERβ (mouse-IgG, 1:800, Sigma-Aldrich), anti-IGF-1R (rabbit-IgG, 1:1000, Santa Cruz), anti-Akt (1:1000, mouse-IgG, Cell Signaling), anti-GSK3-beta (1:500, mouse-IgG, Abcam), anti-GSK3 beta-phospho S9 (GSK3β-p-Ser, 1:500, rabbit-IgG, Abcam), anti-phospho-Akt-Thr308 (p-Akt, 1:1000, rabbit-IgG, Cell Signaling), anti PI3 (1:2000, mouse-IgG, Santa Cruz), anti β-catenin (1:1000, rabbit-IgG, Abcam). .. We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Article Title: CX3CR1 Deficiency Alters Microglial Activation and Reduces Beta-Amyloid Deposition in Two Alzheimer's Disease Mouse Models
    Article Snippet: After overnight incubation at 4°C with primary antibodies, blots were washed with PBS and detected using anti-mouse or anti-rabbit secondary antibodies conjugated to horseradish peroxidase (1:20,000; Jackson ImmunoResearch Laboratories, West Grove, PA) diluted in 5% milk with 0.1% Tween 20. .. To confirm equal protein loading, anti–α-tubulin antibody (mouse monoclonal; 1:5000; Neomarkers, Fremont, CA) or anti–glyceraldehyde 3-phosphate dehydrogenase (GAPDH; mouse monoclonal; 1:20,000; Millipore, Billerica, MA) was used.

    Article Title: Palmitoylethanolamide Ameliorates Hippocampal Damage and Behavioral Dysfunction After Perinatal Asphyxia in the Immature Rat Brain
    Article Snippet: Membranes were blocked with 5% non-fat milk powder and 1% BSA in Tris-buffered saline containing 0.05% Tween 20 and incubated overnight at 4°C with anti-microtubule-associated protein 2 (MAP-2; 1:1,000, polyclonal rabbit-IgG; Abcam), anti-phosphorylated high and medium molecular weight neurofilaments (pNF-H/M; 1:500, monoclonal mouse-IgG; Millipore) and anti-glial fibrillary acidic protein (GFAP; monoclonal mouse-IgG, 1:1,000; Santa Cruz Biotechnology). .. We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1,000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: The membrane was incubated with primary antibodies overnight at 4°C, followed by incubation with secondary antibodies for 2 h at room temperature (RT); both primary and secondary antibodies were prepared in 50% OB buffer. .. The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma).

    Stripping Membranes:

    Article Title: Antibodies to cannabinoid type 1 receptor co-react with stomatin-like protein 2 in mouse brain mitochondria
    Article Snippet: The membranes were subsequently immunoblotted with anti-CB1 (guinea pig; Frontier Science, Japan; 1 : 400), anti-SLP-2 (rabbit; 1 : 200; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (mouse; 1 : 700; Chemicon International, Temecula, CA, USA) for load control. .. For stripping between the immunoblot procedures, membranes were rinsed and incubated in Restore Western Blot Stripping Buffer (Thermo Scientific, Rockford, IL, USA) according to the manufacturer’s instructions.

    BIA-KA:

    Article Title: CX3CR1 Deficiency Alters Microglial Activation and Reduces Beta-Amyloid Deposition in Two Alzheimer's Disease Mouse Models
    Article Snippet: Total protein concentration was determined using the Pierce BCA Protein Assay Kit (Thermo Scientific). .. To confirm equal protein loading, anti–α-tubulin antibody (mouse monoclonal; 1:5000; Neomarkers, Fremont, CA) or anti–glyceraldehyde 3-phosphate dehydrogenase (GAPDH; mouse monoclonal; 1:20,000; Millipore, Billerica, MA) was used.

    Western Blot:

    Article Title: Estradiol Activates PI3K/Akt/GSK3 Pathway Under Chronic Neurodegenerative Conditions Triggered by Perinatal Asphyxia
    Article Snippet: Western blot analysis was carried out using cytosolic fractions separated on 8, 10, or 12.5% SDS-PAGE, in accordance with the molecular weight of the interested protein. .. We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Article Title: CX3CR1 Deficiency Alters Microglial Activation and Reduces Beta-Amyloid Deposition in Two Alzheimer's Disease Mouse Models
    Article Snippet: Paragraph title: Western Blot Analysis ... To confirm equal protein loading, anti–α-tubulin antibody (mouse monoclonal; 1:5000; Neomarkers, Fremont, CA) or anti–glyceraldehyde 3-phosphate dehydrogenase (GAPDH; mouse monoclonal; 1:20,000; Millipore, Billerica, MA) was used.

    Article Title: Palmitoylethanolamide Ameliorates Hippocampal Damage and Behavioral Dysfunction After Perinatal Asphyxia in the Immature Rat Brain
    Article Snippet: Paragraph title: Western blot ... We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1,000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Article Title: Transient activation of AMPK preceding left ventricular pressure overload reduces adverse remodeling and preserves left ventricular function
    Article Snippet: Paragraph title: Western blotting ... Immunoblotting was performed by standard methods with the antibodies indicated [anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, ab2302; MilliporeSigma); anti-p-pyruvate dehydrogenase (ABS203; MilliporeSigma), anti-p-AMPK (sc-33524; Santa Cruz Biotechnology, Dallas, TX, USA), anti-p-eukaryotic elongation factor 2 (eEF2, Thr56, 2331S; Cell Signaling Technology, Danvers, MA, USA), anti-eEF2 (2332S; Cell Signaling Technology), anti-p-Akt (Ser473, 9271S; Cell Signaling Technology), anti-Akt (9272S; Cell Signaling Technology), anti-p-p85 (4228S; Cell Signaling Technology), anti-p-ERK-1/2 (9101S; Cell Signaling Technology), anti-ERK (9102S; Cell Signaling Technology), anti-p-S6 (Ser235/236, 4858S; Cell Signaling Technology), anti-S6 (2317S; Cell Signaling Technology), anti-p70 ribosomal S6 kinase (9202S; Cell Signaling Technology), anti-p-eukaryotic translation initiation factor 4E-binding protein 1 (Ser 65, 9456S; Cell Signaling Technology), anti-IGF1 (sc-9013; Santa Cruz Biotechnology), anti-superoxide dismutase 3 (sc-271170; Santa Cruz Biotechnology), anti-carbonic anhydrase 9 (bs-4029R; Bioss Antibodies, Woburn, MA, USA), anti-p-NOS3 (sc-81510; Santa Cruz Biotechnology), anti-p65 (8242S; Cell Signaling Technology), and anti–forkhead box O 3a (FOXO3a, 12829S; Cell Signaling Technology)].

    Article Title: Antibodies to cannabinoid type 1 receptor co-react with stomatin-like protein 2 in mouse brain mitochondria
    Article Snippet: Paragraph title: Mitochondria/cytosol fractionation and Western blots analysis ... The membranes were subsequently immunoblotted with anti-CB1 (guinea pig; Frontier Science, Japan; 1 : 400), anti-SLP-2 (rabbit; 1 : 200; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (mouse; 1 : 700; Chemicon International, Temecula, CA, USA) for load control.

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma). .. Quantification of fluorescently labeled Western blots was carried out using Image Studio (v3.1) software (LI-COR) and a background subtraction method.

    Article Title: Chondroitin Sulfate Proteoglycan Tenascin-R Regulates Glutamate Uptake by Adult Brain Astrocytes *
    Article Snippet: Paragraph title: Western Blot Analysis ... We used anti-GLAST (1:5000, Cell Signaling Technology), anti-GLT-1 (1:5000, Cell Signaling Technology), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH, 1:5000, mouse monoclonal, Millipore) antibodies.

    Protease Inhibitor:

    Article Title: Transient activation of AMPK preceding left ventricular pressure overload reduces adverse remodeling and preserves left ventricular function
    Article Snippet: Tissue lysates were made from left ventricular sections by homogenizing in 1× heart lysis buffer [25 mM Tris-HCl (pH 7.4), 250 mM NaCl, 5 mM EDTA, 1% Triton X-100, 0.1% SDS, 0.1% sodium deoxycholate, 1 mM DTT, 1× protease inhibitor cocktail (MilliporeSigma, Burlington, MA, USA), 1× phosphatase inhibitor cocktail (F. Hoffman-La Roche)]. .. Immunoblotting was performed by standard methods with the antibodies indicated [anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, ab2302; MilliporeSigma); anti-p-pyruvate dehydrogenase (ABS203; MilliporeSigma), anti-p-AMPK (sc-33524; Santa Cruz Biotechnology, Dallas, TX, USA), anti-p-eukaryotic elongation factor 2 (eEF2, Thr56, 2331S; Cell Signaling Technology, Danvers, MA, USA), anti-eEF2 (2332S; Cell Signaling Technology), anti-p-Akt (Ser473, 9271S; Cell Signaling Technology), anti-Akt (9272S; Cell Signaling Technology), anti-p-p85 (4228S; Cell Signaling Technology), anti-p-ERK-1/2 (9101S; Cell Signaling Technology), anti-ERK (9102S; Cell Signaling Technology), anti-p-S6 (Ser235/236, 4858S; Cell Signaling Technology), anti-S6 (2317S; Cell Signaling Technology), anti-p70 ribosomal S6 kinase (9202S; Cell Signaling Technology), anti-p-eukaryotic translation initiation factor 4E-binding protein 1 (Ser 65, 9456S; Cell Signaling Technology), anti-IGF1 (sc-9013; Santa Cruz Biotechnology), anti-superoxide dismutase 3 (sc-271170; Santa Cruz Biotechnology), anti-carbonic anhydrase 9 (bs-4029R; Bioss Antibodies, Woburn, MA, USA), anti-p-NOS3 (sc-81510; Santa Cruz Biotechnology), anti-p65 (8242S; Cell Signaling Technology), and anti–forkhead box O 3a (FOXO3a, 12829S; Cell Signaling Technology)].

    Article Title: Antibodies to cannabinoid type 1 receptor co-react with stomatin-like protein 2 in mouse brain mitochondria
    Article Snippet: The second supernatants were collected as cytosolic fractions, whereas the pellets were resuspended in 100 μL of mitochondrial extraction buffer mix containing DTT (1 : 1000) and protease inhibitor cocktail (1 : 500; all from Calbiochem, La Jolla, CA, USA) and saved as mitochondrial fractions. .. The membranes were subsequently immunoblotted with anti-CB1 (guinea pig; Frontier Science, Japan; 1 : 400), anti-SLP-2 (rabbit; 1 : 200; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (mouse; 1 : 700; Chemicon International, Temecula, CA, USA) for load control.

    Imaging:

    Article Title: CX3CR1 Deficiency Alters Microglial Activation and Reduces Beta-Amyloid Deposition in Two Alzheimer's Disease Mouse Models
    Article Snippet: To confirm equal protein loading, anti–α-tubulin antibody (mouse monoclonal; 1:5000; Neomarkers, Fremont, CA) or anti–glyceraldehyde 3-phosphate dehydrogenase (GAPDH; mouse monoclonal; 1:20,000; Millipore, Billerica, MA) was used. .. Infrared signals were measured and quantified in an Odyssey infrared imaging system (Li-Cor Biosciences, Lincoln, NE).

    Article Title: Transient activation of AMPK preceding left ventricular pressure overload reduces adverse remodeling and preserves left ventricular function
    Article Snippet: Immunoblotting was performed by standard methods with the antibodies indicated [anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, ab2302; MilliporeSigma); anti-p-pyruvate dehydrogenase (ABS203; MilliporeSigma), anti-p-AMPK (sc-33524; Santa Cruz Biotechnology, Dallas, TX, USA), anti-p-eukaryotic elongation factor 2 (eEF2, Thr56, 2331S; Cell Signaling Technology, Danvers, MA, USA), anti-eEF2 (2332S; Cell Signaling Technology), anti-p-Akt (Ser473, 9271S; Cell Signaling Technology), anti-Akt (9272S; Cell Signaling Technology), anti-p-p85 (4228S; Cell Signaling Technology), anti-p-ERK-1/2 (9101S; Cell Signaling Technology), anti-ERK (9102S; Cell Signaling Technology), anti-p-S6 (Ser235/236, 4858S; Cell Signaling Technology), anti-S6 (2317S; Cell Signaling Technology), anti-p70 ribosomal S6 kinase (9202S; Cell Signaling Technology), anti-p-eukaryotic translation initiation factor 4E-binding protein 1 (Ser 65, 9456S; Cell Signaling Technology), anti-IGF1 (sc-9013; Santa Cruz Biotechnology), anti-superoxide dismutase 3 (sc-271170; Santa Cruz Biotechnology), anti-carbonic anhydrase 9 (bs-4029R; Bioss Antibodies, Woburn, MA, USA), anti-p-NOS3 (sc-81510; Santa Cruz Biotechnology), anti-p65 (8242S; Cell Signaling Technology), and anti–forkhead box O 3a (FOXO3a, 12829S; Cell Signaling Technology)]. .. Secondary antibodies were either fluorophore- or horseradish peroxidase–conjugated, and detection was performed on an Odyssey imaging system (Li-Cor Biosciences, Lincoln, NE, USA).

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma). .. Secondary antibodies were anti-goat (800 nm) or anti-mouse (700 nm), which were used at 1:5,000, prior to imaging fluorescence using a LI-COR Odyssey Sa infrared imaging system.

    Protein Concentration:

    Article Title: Estradiol Activates PI3K/Akt/GSK3 Pathway Under Chronic Neurodegenerative Conditions Triggered by Perinatal Asphyxia
    Article Snippet: Protein concentration was estimated by Bradford technique ( ). .. We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Article Title: CX3CR1 Deficiency Alters Microglial Activation and Reduces Beta-Amyloid Deposition in Two Alzheimer's Disease Mouse Models
    Article Snippet: Total protein concentration was determined using the Pierce BCA Protein Assay Kit (Thermo Scientific). .. To confirm equal protein loading, anti–α-tubulin antibody (mouse monoclonal; 1:5000; Neomarkers, Fremont, CA) or anti–glyceraldehyde 3-phosphate dehydrogenase (GAPDH; mouse monoclonal; 1:20,000; Millipore, Billerica, MA) was used.

    Article Title: Palmitoylethanolamide Ameliorates Hippocampal Damage and Behavioral Dysfunction After Perinatal Asphyxia in the Immature Rat Brain
    Article Snippet: The supernatants were analyzed for total protein concentration using Bradford solution (Bio-Rad, Richmond CA, USA) in 96-well plates using bovine serum albumin (BSA) as standard. .. We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1,000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: Cells were washed twice in PBS, lysed in 1× Glasgow lysis buffer (GLB; 1% [vol/vol] Triton X-100, 120 mM KCl, 30 mM NaCl, 5 mM MgCl2 , 10% [vol/vol] glycerol, 10 mM PIPES [piperazine-N ,N ′-bis(2-ethanesulfonic acid)]-NaOH, pH 7.2, with protease and phosphatase inhibitors), and clarified by centrifugation at 2,800 × g for 5 min at 4°C, before determining and normalizing the protein concentration by a bicinchoninic acid assay (Pierce). .. The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma).

    Sonication:

    Article Title: CX3CR1 Deficiency Alters Microglial Activation and Reduces Beta-Amyloid Deposition in Two Alzheimer's Disease Mouse Models
    Article Snippet: Total brain homogenates were subsequently sonicated to shear the DNA and centrifuged to remove nuclei and cellular debris. .. To confirm equal protein loading, anti–α-tubulin antibody (mouse monoclonal; 1:5000; Neomarkers, Fremont, CA) or anti–glyceraldehyde 3-phosphate dehydrogenase (GAPDH; mouse monoclonal; 1:20,000; Millipore, Billerica, MA) was used.

    Molecular Weight:

    Article Title: Estradiol Activates PI3K/Akt/GSK3 Pathway Under Chronic Neurodegenerative Conditions Triggered by Perinatal Asphyxia
    Article Snippet: Western blot analysis was carried out using cytosolic fractions separated on 8, 10, or 12.5% SDS-PAGE, in accordance with the molecular weight of the interested protein. .. We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Article Title: Palmitoylethanolamide Ameliorates Hippocampal Damage and Behavioral Dysfunction After Perinatal Asphyxia in the Immature Rat Brain
    Article Snippet: Membranes were blocked with 5% non-fat milk powder and 1% BSA in Tris-buffered saline containing 0.05% Tween 20 and incubated overnight at 4°C with anti-microtubule-associated protein 2 (MAP-2; 1:1,000, polyclonal rabbit-IgG; Abcam), anti-phosphorylated high and medium molecular weight neurofilaments (pNF-H/M; 1:500, monoclonal mouse-IgG; Millipore) and anti-glial fibrillary acidic protein (GFAP; monoclonal mouse-IgG, 1:1,000; Santa Cruz Biotechnology). .. We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1,000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Fluorescence:

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma). .. Secondary antibodies were anti-goat (800 nm) or anti-mouse (700 nm), which were used at 1:5,000, prior to imaging fluorescence using a LI-COR Odyssey Sa infrared imaging system.

    Immunodetection:

    Article Title: Chondroitin Sulfate Proteoglycan Tenascin-R Regulates Glutamate Uptake by Adult Brain Astrocytes *
    Article Snippet: We used anti-GLAST (1:5000, Cell Signaling Technology), anti-GLT-1 (1:5000, Cell Signaling Technology), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH, 1:5000, mouse monoclonal, Millipore) antibodies. .. Immunodetection was performed using the ECL Prime or Select Western blotting detection System (GE Healthcare) with horseradish peroxidase-conjugated antibodies against mouse or rabbit IgG (1:10,000, Cell Signaling Technology) according to the manufacturer's instructions.

    Labeling:

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma). .. Quantification of fluorescently labeled Western blots was carried out using Image Studio (v3.1) software (LI-COR) and a background subtraction method.

    Mouse Assay:

    Article Title: Antibodies to cannabinoid type 1 receptor co-react with stomatin-like protein 2 in mouse brain mitochondria
    Article Snippet: Either single embryo brain or one adult cerebral hemisphere from adult mice were homogenized in an ice-cold tissue grinder with 0.5–1.0 mL cytosol extraction buffer mix containing dithiothreitol (DTT; 1 : 1000) and protease inhibitor cocktail (1 : 500; all from Calbiochem, La Jolla, CA, USA). .. The membranes were subsequently immunoblotted with anti-CB1 (guinea pig; Frontier Science, Japan; 1 : 400), anti-SLP-2 (rabbit; 1 : 200; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (mouse; 1 : 700; Chemicon International, Temecula, CA, USA) for load control.

    Stripping:

    Article Title: Antibodies to cannabinoid type 1 receptor co-react with stomatin-like protein 2 in mouse brain mitochondria
    Article Snippet: The membranes were subsequently immunoblotted with anti-CB1 (guinea pig; Frontier Science, Japan; 1 : 400), anti-SLP-2 (rabbit; 1 : 200; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (mouse; 1 : 700; Chemicon International, Temecula, CA, USA) for load control. .. For stripping between the immunoblot procedures, membranes were rinsed and incubated in Restore Western Blot Stripping Buffer (Thermo Scientific, Rockford, IL, USA) according to the manufacturer’s instructions.

    Staining:

    Article Title: Transient activation of AMPK preceding left ventricular pressure overload reduces adverse remodeling and preserves left ventricular function
    Article Snippet: Immunoblotting was performed by standard methods with the antibodies indicated [anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, ab2302; MilliporeSigma); anti-p-pyruvate dehydrogenase (ABS203; MilliporeSigma), anti-p-AMPK (sc-33524; Santa Cruz Biotechnology, Dallas, TX, USA), anti-p-eukaryotic elongation factor 2 (eEF2, Thr56, 2331S; Cell Signaling Technology, Danvers, MA, USA), anti-eEF2 (2332S; Cell Signaling Technology), anti-p-Akt (Ser473, 9271S; Cell Signaling Technology), anti-Akt (9272S; Cell Signaling Technology), anti-p-p85 (4228S; Cell Signaling Technology), anti-p-ERK-1/2 (9101S; Cell Signaling Technology), anti-ERK (9102S; Cell Signaling Technology), anti-p-S6 (Ser235/236, 4858S; Cell Signaling Technology), anti-S6 (2317S; Cell Signaling Technology), anti-p70 ribosomal S6 kinase (9202S; Cell Signaling Technology), anti-p-eukaryotic translation initiation factor 4E-binding protein 1 (Ser 65, 9456S; Cell Signaling Technology), anti-IGF1 (sc-9013; Santa Cruz Biotechnology), anti-superoxide dismutase 3 (sc-271170; Santa Cruz Biotechnology), anti-carbonic anhydrase 9 (bs-4029R; Bioss Antibodies, Woburn, MA, USA), anti-p-NOS3 (sc-81510; Santa Cruz Biotechnology), anti-p65 (8242S; Cell Signaling Technology), and anti–forkhead box O 3a (FOXO3a, 12829S; Cell Signaling Technology)]. .. Total protein stain was performed with Revert Total Protein Stain, according to manufacturer’s instructions (Li-Cor Biosciences).

    SDS Page:

    Article Title: Estradiol Activates PI3K/Akt/GSK3 Pathway Under Chronic Neurodegenerative Conditions Triggered by Perinatal Asphyxia
    Article Snippet: Western blot analysis was carried out using cytosolic fractions separated on 8, 10, or 12.5% SDS-PAGE, in accordance with the molecular weight of the interested protein. .. We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Article Title: Palmitoylethanolamide Ameliorates Hippocampal Damage and Behavioral Dysfunction After Perinatal Asphyxia in the Immature Rat Brain
    Article Snippet: The samples were subjected to SDS-PAGE using the Mini-protein II cell (Bio-Rad, Richmond CA, USA) with precast 4–20% Precise gels (Bio-Rad, Richmond CA, USA). .. We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1,000, rabbit-IgG, Sigma-Aldrich) as loading control.

    Article Title: Transient activation of AMPK preceding left ventricular pressure overload reduces adverse remodeling and preserves left ventricular function
    Article Snippet: SDS-PAGE and protein transfer to PVDF were performed following standard procedures. .. Immunoblotting was performed by standard methods with the antibodies indicated [anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, ab2302; MilliporeSigma); anti-p-pyruvate dehydrogenase (ABS203; MilliporeSigma), anti-p-AMPK (sc-33524; Santa Cruz Biotechnology, Dallas, TX, USA), anti-p-eukaryotic elongation factor 2 (eEF2, Thr56, 2331S; Cell Signaling Technology, Danvers, MA, USA), anti-eEF2 (2332S; Cell Signaling Technology), anti-p-Akt (Ser473, 9271S; Cell Signaling Technology), anti-Akt (9272S; Cell Signaling Technology), anti-p-p85 (4228S; Cell Signaling Technology), anti-p-ERK-1/2 (9101S; Cell Signaling Technology), anti-ERK (9102S; Cell Signaling Technology), anti-p-S6 (Ser235/236, 4858S; Cell Signaling Technology), anti-S6 (2317S; Cell Signaling Technology), anti-p70 ribosomal S6 kinase (9202S; Cell Signaling Technology), anti-p-eukaryotic translation initiation factor 4E-binding protein 1 (Ser 65, 9456S; Cell Signaling Technology), anti-IGF1 (sc-9013; Santa Cruz Biotechnology), anti-superoxide dismutase 3 (sc-271170; Santa Cruz Biotechnology), anti-carbonic anhydrase 9 (bs-4029R; Bioss Antibodies, Woburn, MA, USA), anti-p-NOS3 (sc-81510; Santa Cruz Biotechnology), anti-p65 (8242S; Cell Signaling Technology), and anti–forkhead box O 3a (FOXO3a, 12829S; Cell Signaling Technology)].

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: Following separation by SDS-PAGE, proteins were transferred to a polyvinylidene difluoride (PVDF) membrane and blocked in 50% (vol/vol) Odyssey blocking (OB) buffer (LI-COR) in PBS. .. The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma).

    Software:

    Article Title: Estradiol Activates PI3K/Akt/GSK3 Pathway Under Chronic Neurodegenerative Conditions Triggered by Perinatal Asphyxia
    Article Snippet: We used anti glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:1000, rabbit-IgG, Sigma-Aldrich) as loading control. .. Films were scanned and the optical density of protein bands was quantified using Gel Pro Analyzer software 3.1.00.00 (Media Cybernetics).

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma). .. Quantification of fluorescently labeled Western blots was carried out using Image Studio (v3.1) software (LI-COR) and a background subtraction method.

    Article Title: Chondroitin Sulfate Proteoglycan Tenascin-R Regulates Glutamate Uptake by Adult Brain Astrocytes *
    Article Snippet: We used anti-GLAST (1:5000, Cell Signaling Technology), anti-GLT-1 (1:5000, Cell Signaling Technology), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH, 1:5000, mouse monoclonal, Millipore) antibodies. .. Semi-quantification was done by densitometry of blot signals using ImageJ software.

    Acid Assay:

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: Cells were washed twice in PBS, lysed in 1× Glasgow lysis buffer (GLB; 1% [vol/vol] Triton X-100, 120 mM KCl, 30 mM NaCl, 5 mM MgCl2 , 10% [vol/vol] glycerol, 10 mM PIPES [piperazine-N ,N ′-bis(2-ethanesulfonic acid)]-NaOH, pH 7.2, with protease and phosphatase inhibitors), and clarified by centrifugation at 2,800 × g for 5 min at 4°C, before determining and normalizing the protein concentration by a bicinchoninic acid assay (Pierce). .. The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma).

    Fractionation:

    Article Title: Antibodies to cannabinoid type 1 receptor co-react with stomatin-like protein 2 in mouse brain mitochondria
    Article Snippet: Paragraph title: Mitochondria/cytosol fractionation and Western blots analysis ... The membranes were subsequently immunoblotted with anti-CB1 (guinea pig; Frontier Science, Japan; 1 : 400), anti-SLP-2 (rabbit; 1 : 200; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (mouse; 1 : 700; Chemicon International, Temecula, CA, USA) for load control.

    Lysis:

    Article Title: Transient activation of AMPK preceding left ventricular pressure overload reduces adverse remodeling and preserves left ventricular function
    Article Snippet: Tissue lysates were made from left ventricular sections by homogenizing in 1× heart lysis buffer [25 mM Tris-HCl (pH 7.4), 250 mM NaCl, 5 mM EDTA, 1% Triton X-100, 0.1% SDS, 0.1% sodium deoxycholate, 1 mM DTT, 1× protease inhibitor cocktail (MilliporeSigma, Burlington, MA, USA), 1× phosphatase inhibitor cocktail (F. Hoffman-La Roche)]. .. Immunoblotting was performed by standard methods with the antibodies indicated [anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, ab2302; MilliporeSigma); anti-p-pyruvate dehydrogenase (ABS203; MilliporeSigma), anti-p-AMPK (sc-33524; Santa Cruz Biotechnology, Dallas, TX, USA), anti-p-eukaryotic elongation factor 2 (eEF2, Thr56, 2331S; Cell Signaling Technology, Danvers, MA, USA), anti-eEF2 (2332S; Cell Signaling Technology), anti-p-Akt (Ser473, 9271S; Cell Signaling Technology), anti-Akt (9272S; Cell Signaling Technology), anti-p-p85 (4228S; Cell Signaling Technology), anti-p-ERK-1/2 (9101S; Cell Signaling Technology), anti-ERK (9102S; Cell Signaling Technology), anti-p-S6 (Ser235/236, 4858S; Cell Signaling Technology), anti-S6 (2317S; Cell Signaling Technology), anti-p70 ribosomal S6 kinase (9202S; Cell Signaling Technology), anti-p-eukaryotic translation initiation factor 4E-binding protein 1 (Ser 65, 9456S; Cell Signaling Technology), anti-IGF1 (sc-9013; Santa Cruz Biotechnology), anti-superoxide dismutase 3 (sc-271170; Santa Cruz Biotechnology), anti-carbonic anhydrase 9 (bs-4029R; Bioss Antibodies, Woburn, MA, USA), anti-p-NOS3 (sc-81510; Santa Cruz Biotechnology), anti-p65 (8242S; Cell Signaling Technology), and anti–forkhead box O 3a (FOXO3a, 12829S; Cell Signaling Technology)].

    Article Title: Serine Phosphorylation of the Hepatitis C Virus NS5A Protein Controls the Establishment of Replication Complexes
    Article Snippet: Cells were washed twice in PBS, lysed in 1× Glasgow lysis buffer (GLB; 1% [vol/vol] Triton X-100, 120 mM KCl, 30 mM NaCl, 5 mM MgCl2 , 10% [vol/vol] glycerol, 10 mM PIPES [piperazine-N ,N ′-bis(2-ethanesulfonic acid)]-NaOH, pH 7.2, with protease and phosphatase inhibitors), and clarified by centrifugation at 2,800 × g for 5 min at 4°C, before determining and normalizing the protein concentration by a bicinchoninic acid assay (Pierce). .. The primary antibodies used were anti-NS5A (sheep, 1:5,000) , anti-NS3 (sheep, 1:2,000; prepared in-house), and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH; mouse, 1:20,000; Sigma).

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  • 90
    Millipore mouse anti bax monoclonal
    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of <t>pro-caspase-8/9/3</t> (A) and <t>Bcl-2/Bax</t> (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P
    Mouse Anti Bax Monoclonal, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti bax monoclonal/product/Millipore
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    92
    Millipore mouse monoclonal anti glyceraldehyde 3 phosphate dehydrogenase gapdh antibody
    Sumoylation of FOXP1. ( A ) Representative immunoblotting of Foxp1 in the mouse neocortex during development. ( Right panel) Quantification of SUMO–Foxp1 immunoblotting. Immunoblots were first normalized to <t>glyceraldehyde-3-phosphate</t> dehydrogenase <t>(GAPDH)</t> at each time point and then subsequently normalized to nonsumoylated Foxp1 levels at embryonic day 15.5 (E15.5). Data are represented as means (±SEM). n = 3 per condition. ( B ) Endogenous coimmunoprecipitation of Foxp1 and SUMO-1 in the mouse neocortex at P0. ( C ) Schematic of FOXP1 protein showing the location of K636. (PolyQ) Polyglutamine motif; (ZF) zinc finger; (LZ) leucine zipper. ( D ) K636 is conserved across species. ( E ) Immunoblotting for Flag-tagged FOXP1 in 293T cells. Lysates were treated with 1 mM H 2 O 2 for 1 h ( left panel) or 100 µM ginkgolic acid for 6 h ( right panel) in the presence or absence of NEM. (FOXP1 WT) Flag-tagged wild-type FOXP1. The asterisk indicates a nonspecific band of the SUMO-1 antibody. ( F ) Immunoblot of immunoprecipitated wild-type Flag-tagged FOXP1 or Flag-tagged FOXP1 KR.
    Mouse Monoclonal Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti glyceraldehyde 3 phosphate dehydrogenase gapdh antibody/product/Millipore
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    mouse monoclonal anti glyceraldehyde 3 phosphate dehydrogenase gapdh antibody - by Bioz Stars, 2020-04
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    90
    Millipore rabbit anti bax
    Effects of coffee oil-algae oil nanoemulsions on protein expressions of <t>cyclin</t> B, CDK2, cyclin A, and CDK1 ( A ), p53 and p21 ( B ), and <t>Bax,</t> Bcl-2, cytochrome C ( C ). Notes: Control cells are incubated with medium only. Results are presented as mean ± standard deviation of triplicate determinations. Data with different letters (A–C) are significantly different at p
    Rabbit Anti Bax, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti bax/product/Millipore
    Average 90 stars, based on 9 article reviews
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    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Expressing, Cell Culture, Transfection, Western Blot, Positive Control

    Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Activation Assay, Expressing, Cell Culture, Transfection, Western Blot

    Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Expressing, Western Blot

    Sumoylation of FOXP1. ( A ) Representative immunoblotting of Foxp1 in the mouse neocortex during development. ( Right panel) Quantification of SUMO–Foxp1 immunoblotting. Immunoblots were first normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) at each time point and then subsequently normalized to nonsumoylated Foxp1 levels at embryonic day 15.5 (E15.5). Data are represented as means (±SEM). n = 3 per condition. ( B ) Endogenous coimmunoprecipitation of Foxp1 and SUMO-1 in the mouse neocortex at P0. ( C ) Schematic of FOXP1 protein showing the location of K636. (PolyQ) Polyglutamine motif; (ZF) zinc finger; (LZ) leucine zipper. ( D ) K636 is conserved across species. ( E ) Immunoblotting for Flag-tagged FOXP1 in 293T cells. Lysates were treated with 1 mM H 2 O 2 for 1 h ( left panel) or 100 µM ginkgolic acid for 6 h ( right panel) in the presence or absence of NEM. (FOXP1 WT) Flag-tagged wild-type FOXP1. The asterisk indicates a nonspecific band of the SUMO-1 antibody. ( F ) Immunoblot of immunoprecipitated wild-type Flag-tagged FOXP1 or Flag-tagged FOXP1 KR.

    Journal: Genes & Development

    Article Title: Foxp1 regulation of neonatal vocalizations via cortical development

    doi: 10.1101/gad.305037.117

    Figure Lengend Snippet: Sumoylation of FOXP1. ( A ) Representative immunoblotting of Foxp1 in the mouse neocortex during development. ( Right panel) Quantification of SUMO–Foxp1 immunoblotting. Immunoblots were first normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) at each time point and then subsequently normalized to nonsumoylated Foxp1 levels at embryonic day 15.5 (E15.5). Data are represented as means (±SEM). n = 3 per condition. ( B ) Endogenous coimmunoprecipitation of Foxp1 and SUMO-1 in the mouse neocortex at P0. ( C ) Schematic of FOXP1 protein showing the location of K636. (PolyQ) Polyglutamine motif; (ZF) zinc finger; (LZ) leucine zipper. ( D ) K636 is conserved across species. ( E ) Immunoblotting for Flag-tagged FOXP1 in 293T cells. Lysates were treated with 1 mM H 2 O 2 for 1 h ( left panel) or 100 µM ginkgolic acid for 6 h ( right panel) in the presence or absence of NEM. (FOXP1 WT) Flag-tagged wild-type FOXP1. The asterisk indicates a nonspecific band of the SUMO-1 antibody. ( F ) Immunoblot of immunoprecipitated wild-type Flag-tagged FOXP1 or Flag-tagged FOXP1 KR.

    Article Snippet: The following antibodies were used: mouse monoclonal anti-SUMO-1 (D-11) antibody (Santa Cruz Biotechnology, sc-5308), rabbit polyclonal anti-FOXP1 antibody , mouse monoclonal anti-FOXP1 (JC12) antibody (Abcam, ab32010), goat polyclonal anti-FOXP2 (N-16) antibody (Santa Cruz Biotechnology, sc-21068), mouse monoclonal anti-Flag M2 antibody (Sigma-Aldrich, F1804), mouse monoclonal anti-V5 antibody (Invitrogen, R960-25), goat polyclonal anti-GFP antibody (Rockland Immunochemicals, 600-101-215), chick polyclonal anti-GFP antibody (Aves Laboratories, GFP-1010), rabbit monoclonal anti-SUMO-2/3 (18H8) antibody (Cell Signaling Technology, 4971), rabbit polyclonal anti-PIAS2 antibody (Abcam, ab155556), rabbit polyclonal anti-PIAS3 (H-169) antibody (Santa Cruz Biotechnology, sc-14017), rabbit polyclonal anti-MAP2 antibody (Chemicon, AB5622), mouse monoclonal anti-CtBP (E-12) antibody (Santa Cruz Biotechnology, sc-17759), rabbit polyclonal anti-CDP (CUX1: M-222) antibody (Santa Cruz Biotechnology, sc-13024), rat anti-CTIP2 (Abcam, ab18465), rabbit polyclonal anti-HDAC1 antibody (Abcam, ab19845), mouse monoclonal anti-HDAC1 (10E2) antibody (Cell Signaling Technology, 5256), mouse monoclonal anti-HDAC2 (3F3) antibody (Cell Signaling Technology, 5113), rabbit monoclonal anti-MTA1 (D40D1) XP antibody (Cell Signaling Technology, 5647), rabbit polyclonal anti-MTA2 (H-170) antibody (Santa Cruz Biotechnology, sc-28731), rabbit polyclonal anti-p66β (GATAD2B) antibody (Novus Biologicals, NBP1-87358), mouse monoclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (Millipore, MAB374), mouse (G3A1) mAb IgG1 isotype control (Cell Signaling Technology, 5415), normal rabbit IgG (Cell Signaling Technology, 2729), and normal goat IgG (Santa Cruz Biotechnology, sc-2028).

    Techniques: Western Blot, Immunoprecipitation

    Effects of coffee oil-algae oil nanoemulsions on protein expressions of cyclin B, CDK2, cyclin A, and CDK1 ( A ), p53 and p21 ( B ), and Bax, Bcl-2, cytochrome C ( C ). Notes: Control cells are incubated with medium only. Results are presented as mean ± standard deviation of triplicate determinations. Data with different letters (A–C) are significantly different at p

    Journal: International Journal of Nanomedicine

    Article Title: Preparation of coffee oil-algae oil-based nanoemulsions and the study of their inhibition effect on UVA-induced skin damage in mice and melanoma cell growth

    doi: 10.2147/IJN.S144705

    Figure Lengend Snippet: Effects of coffee oil-algae oil nanoemulsions on protein expressions of cyclin B, CDK2, cyclin A, and CDK1 ( A ), p53 and p21 ( B ), and Bax, Bcl-2, cytochrome C ( C ). Notes: Control cells are incubated with medium only. Results are presented as mean ± standard deviation of triplicate determinations. Data with different letters (A–C) are significantly different at p

    Article Snippet: The primary antibodies include mouse monoclonal anti-α-tubulin antibody (Sigma–Aldrich Co.), mouse anti-cyclin A and rabbit anti-Bax (EMD Millipore), mouse anti-CDK2, mouse anti-cytochrome C, mouse anti-p21, mouse anti-cyclin B, and mouse anti-Bcl-2 (BD Biosciences), as well as anti-cdc2 (CDK1) and mouse anti-p53 (Novus Biologicals Co., Littleton, CO, USA).

    Techniques: Incubation, Standard Deviation