mouse anti glyceraldehyde 3 phosphate dehydrogenase gapdh  (Millipore)


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    Structured Review

    Millipore mouse anti glyceraldehyde 3 phosphate dehydrogenase gapdh
    Mouse Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti glyceraldehyde 3 phosphate dehydrogenase gapdh/product/Millipore
    Average 96 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    mouse anti glyceraldehyde 3 phosphate dehydrogenase gapdh - by Bioz Stars, 2020-03
    96/100 stars

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    Related Articles

    Transduction:

    Article Title: Nitric Oxide Induces the Progression of Abdominal Aortic Aneurysms through the Matrix Metalloproteinase Inducer EMMPRIN
    Article Snippet: .. Primary antibodies were obtained as follows: rabbit anti-MMP-13 from Calbiochem; goat-anti-human-MMP-13 and rabbit anti-inducible NO synthase (iNOS) from Santa Cruz Biotechnology (Santa Cruz, CA); rabbit anti-MMP-2, mouse anti-MMP-9, rabbit anti-MMP-12, and mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Chemicon (Millipore, Iberica, Spain); anti-FLAG, and goat-anti-EMMPRIN from Sigma-Aldrich; anti-ICAM-2, anti-von Willebrand factor, and anti-Mac-3 from Transduction Laboratories (BD Biosciences). .. Rat-anti-mouse polymorphonuclear from Serotec (Oxford, UK).

    Article Title: Vimentin Regulates Scribble Activity by Protecting It from Proteasomal Degradation
    Article Snippet: .. Monoclonal antibodies were mouse anti-GM130 clone 35 and anti-β-Catenin clone 14 (BD Biosciences Transduction Laboratories, Lexington, KY), mouse anti-vimentin clone V9 and anti-α-tubulin clone GTU-88 (Sigma-Aldrich), rat anti-HA clone 3F10 (Roche Diagnostics, Indianapolis, IN), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) clone 6C5 (Millipore Bioscience Research Reagents, Temecula, CA), mouse anti-keratin 18 clone C-04 (Abcam), and mouse anti-keratin 18 clone LDK18, a gift from Birgit E. Lane (Institute of Medical Biology, Immunos, Singapore). .. Control antibodies used in immunoprecipitations were normal mouse and goat immunoglobulin G (IgG) (Santa Cruz Biotechnology) Secondary antibodies used for immunofluorescence were donkey anti-mouse IgG Alexa Fluor 488 and 594, anti-goat IgG Alexa Fluor 488 and 594, and anti-rabbit IgG Alexa Fluor 488 (Invitrogen) and donkey anti-mouse IgG 7-amino-4-methylcoumarin-3-acetic acid (Jackson ImmunoResearch Laboratories, West Grove, PA).

    Blocking Assay:

    Article Title: Dynamic Regulation of Oct1 during Mitosis by Phosphorylation and Ubiquitination
    Article Snippet: The antibody was purified in two steps, first by blocking with unphosphorylated peptide to remove non-phospho-specific antibodies, then by affinity purification using the phosphorylated epitope. .. Mouse anti-pS10-H3 was obtained from Cell Signaling, and mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Chemicon.

    Article Title: Localization and expression of GABA transporters in the suprachiasmatic nucleus
    Article Snippet: For a loading control a mouse anti-Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) purified monoclonal antibody (MAB374, Millipore, Temecula, CA, 1:8000) was applied followed by rabbit polyclonal anti-mouse IgG-HRP conjugated secondary antibody (AP160P, Millipore, 1:200,000). .. For GAT3 detection, the blots were blocked and incubated overnight in Odyssey blocking buffer containing a rabbit anti-GAT3 polyclonal antibody (AB1574, Millipore, 1:1000).

    Article Title: The glutamate transporter, GLAST, participates in a macromolecular complex that supports glutamate metabolism
    Article Snippet: After blocking for 1 hr at 25°C in TBS (50 mM Tris, pH 8.0, 150 mM NaCl) containing 5% nonfat dry milk, membranes were probed with the appropriate primary antibody. .. Mouse anti-hexokinase (1:1000), mouse anti-α1 subunit of the Na+ /K+ ATPase (1:2000), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:200), and rabbit anti-α2 subunit of the Na+ /K+ ATPase (1:1000) were from Millipore (Temecula, CA).

    Article Title: GABA transporters regulate tonic and synaptic GABAA receptor-mediated currents in the suprachiasmatic nucleus neurons
    Article Snippet: The loading control was mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) purified monoclonal antibody (MAB374; Millipore, Temecula, CA; 1:8,000). .. For GAT3 detection, the blots were blocked and incubated overnight in Odyssey blocking buffer containing a rabbit anti-GAT3 polyclonal antibody (catalog no. AB1574, amino acids 607–627, COOH terminus; Millipore; 1:1,000).

    Electrophoresis:

    Article Title: Monomethyl Auristatin E Phosphate Inhibits Human Prostate Cancer Growth
    Article Snippet: Approximately 50 μg of protein per sample was loaded onto a 10% sodium dodecyl sulfate -polyacrylamide gel and subjected to electrophoresis for 3 hours at 200 volts. .. Antibodies used include the following: mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and rabbit anti-cleaved poly ADP ribose polymerase (PARP) (Millipore, Billerica, MA).

    Incubation:

    Article Title: Glucose and glutamine handling in the Sertoli cells of transgenic rats overexpressing regucalcin: plasticity towards lactate production
    Article Snippet: Membranes were incubated overnight at 4 °C with rabbit anti-GLUT1 (1:500, CBL242, Millipore, MA, USA), rabbit anti-GLUT2 (1:500, sc-9117, Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-GLUT3 (1:1000, sc-30107, Santa Cruz Biotechnology), rabbit anti- PFK1 (1:500, sc-67028, Santa Cruz Biotechnology), rabbit anti-MCT4 (1:1000, sc-50329, Santa Cruz Biotechnology), rabbit anti-LDHA (1:10000, Ab52488, Abcam, Cambridge, MA, USA), mouse anti-ALT (1:200, sc-374501, Santa Cruz Biotechnology), rabbit anti-GLS (1:1000, ab93434, Abcam) and rabbit anti-ASCT2 (1:1000, V501, Cell signalling technology, Danvers, MA, USA) primary antibodies. .. A mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:10000, AB2302, Millipore, Darmstadt, Germany) antibody was used for protein loading control in all WB analyses.

    Article Title: Localization and expression of GABA transporters in the suprachiasmatic nucleus
    Article Snippet: For the ECL detection, the blots were blocked in 5% non-fat milk/TBST and incubated with a rabbit anti-GAT1 polyclonal antibody (AGT-001, Alomone Labs Ltd., Jerusalem, Israel, 1:500) overnight at 4°C in the same buffer with agitation. .. For a loading control a mouse anti-Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) purified monoclonal antibody (MAB374, Millipore, Temecula, CA, 1:8000) was applied followed by rabbit polyclonal anti-mouse IgG-HRP conjugated secondary antibody (AP160P, Millipore, 1:200,000).

    Article Title: The glutamate transporter, GLAST, participates in a macromolecular complex that supports glutamate metabolism
    Article Snippet: Mouse anti-hexokinase (1:1000), mouse anti-α1 subunit of the Na+ /K+ ATPase (1:2000), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:200), and rabbit anti-α2 subunit of the Na+ /K+ ATPase (1:1000) were from Millipore (Temecula, CA). .. Membranes were washed in TBS-T (50 mM Tris, pH 8.0, 150 mM NaCl, 0.1% Tween 20) and then incubated with fluorescently conjugated anti-rabbit, anti-mouse, or anti-goat antibodies (1:10,000 LiCor Biosciences, Lincoln, NE).

    Article Title: Monomethyl Auristatin E Phosphate Inhibits Human Prostate Cancer Growth
    Article Snippet: The membrane was then probed with primary antibody overnight at 4°C and then incubated with IRDye® 800CW and IRDye® 680RD secondary antibodies (LI-COR Biosciences, Lincoln, NE). .. Antibodies used include the following: mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and rabbit anti-cleaved poly ADP ribose polymerase (PARP) (Millipore, Billerica, MA).

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences. .. For immunofluorescence, cells were grown on LabTek 8-well chamber slides, and fixed with 4% paraformaldehyde, followed by treatment with ice-cold methanol for 10 min. After permeabilization with 0.5% Triton X-100 in PBS for 3 min, cells were blocked with 10% normal goat serum and incubated with indicated primary antibodies.

    Article Title: GABA transporters regulate tonic and synaptic GABAA receptor-mediated currents in the suprachiasmatic nucleus neurons
    Article Snippet: For the enhanced chemiluminescence detection, the blots were blocked in 5% non-fat milk in TBST (Tris-buffered saline-Tween 20) and incubated with a rabbit anti-GAT1 (extracellular domain) polyclonal antibody (AGT-001; Alomone Laboratories, Jerusalem, Israel 1:500) overnight at 4°C in the same buffer with agitation. .. The loading control was mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) purified monoclonal antibody (MAB374; Millipore, Temecula, CA; 1:8,000).

    Activity Assay:

    Article Title: Nitric Oxide Induces the Progression of Abdominal Aortic Aneurysms through the Matrix Metalloproteinase Inducer EMMPRIN
    Article Snippet: Porcine elastase (specific activity 5 U/mg; E1250) was from Sigma-Aldrich. .. Primary antibodies were obtained as follows: rabbit anti-MMP-13 from Calbiochem; goat-anti-human-MMP-13 and rabbit anti-inducible NO synthase (iNOS) from Santa Cruz Biotechnology (Santa Cruz, CA); rabbit anti-MMP-2, mouse anti-MMP-9, rabbit anti-MMP-12, and mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Chemicon (Millipore, Iberica, Spain); anti-FLAG, and goat-anti-EMMPRIN from Sigma-Aldrich; anti-ICAM-2, anti-von Willebrand factor, and anti-Mac-3 from Transduction Laboratories (BD Biosciences).

    Expressing:

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: Western blotting and immunofluorescence Whole cell extracts were used to detect the expression of EMT markers by western blotting. .. Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences.

    Modification:

    Article Title: c-Cbl and Cbl-b Act Redundantly to Protect Osteoclasts from Apoptosis and to Displace HDAC6 from ?-Tubulin, Stabilizing Microtubules and Podosomes
    Article Snippet: Minimal essential medium, alpha modification (α-MEM), fetal bovine serum (FBS), fluorescein-conjugated secondary antibodies, and TO-PRO-3 were purchased from Invitrogen (Carlsbad, CA). .. Mouse anti-β-tubulin, mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), mouse anti-HDAC6 and mouse anti-actin antibodies were obtained from Millipore Bioscience Research Reagents (Temecula, CA).

    Western Blot:

    Article Title: A20 Promotes Liver Regeneration by Decreasing SOCS3 Expression to Enhance IL-6/STAT3 Proliferative Signals
    Article Snippet: .. Samples were analyzed by Western Blot (WB) using the following primary antibodies: rabbit anti-STAT3, rabbit anti-IκBα, mouse anti-βactin, (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), rabbit anti-phospho-STAT3 (P-STAT3 Tyr705) (Cell Signaling Technology, Danvers, MA), chicken anti-TNFAIP3 (A20) (Abcam Inc., Cambridge, MA), mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Calbiochem/EMD Biosciences Inc., San Diego, CA), anti-hemagglutinin (HA) (Roche Applied Science, Indianapolis, IN); and secondary antibodies (Thermo Scientific, Rockford, IL). .. Immunoblots were scanned and band intensity quantified by densitometry using ImageJ 1.41 (NIH, Bethesda, MD).

    Article Title: Glucose and glutamine handling in the Sertoli cells of transgenic rats overexpressing regucalcin: plasticity towards lactate production
    Article Snippet: .. A mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:10000, AB2302, Millipore, Darmstadt, Germany) antibody was used for protein loading control in all WB analyses. .. Goat anti-rabbit IgG-HRP (1:5000, NIF1317; Santa Cruz Biotechnology) or goat anti-mouse IgG + IgM-HRP (1:5000, Santa Cruz Biotechnology) were used as secondary antibodies.

    Article Title: Localization and expression of GABA transporters in the suprachiasmatic nucleus
    Article Snippet: Paragraph title: Western blot procedures and data analysis ... For a loading control a mouse anti-Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) purified monoclonal antibody (MAB374, Millipore, Temecula, CA, 1:8000) was applied followed by rabbit polyclonal anti-mouse IgG-HRP conjugated secondary antibody (AP160P, Millipore, 1:200,000).

    Article Title: Vimentin Regulates Scribble Activity by Protecting It from Proteasomal Degradation
    Article Snippet: Monoclonal antibodies were mouse anti-GM130 clone 35 and anti-β-Catenin clone 14 (BD Biosciences Transduction Laboratories, Lexington, KY), mouse anti-vimentin clone V9 and anti-α-tubulin clone GTU-88 (Sigma-Aldrich), rat anti-HA clone 3F10 (Roche Diagnostics, Indianapolis, IN), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) clone 6C5 (Millipore Bioscience Research Reagents, Temecula, CA), mouse anti-keratin 18 clone C-04 (Abcam), and mouse anti-keratin 18 clone LDK18, a gift from Birgit E. Lane (Institute of Medical Biology, Immunos, Singapore). .. For Western blots, horseradish peroxidase (HRP)-coupled goat antibodies to mouse, rabbit (Bio-Rad Laboratories, Hercules, CA), or rat IgG (Pierce Chemical.

    Article Title: The glutamate transporter, GLAST, participates in a macromolecular complex that supports glutamate metabolism
    Article Snippet: Paragraph title: 2.2. Western Blotting ... Mouse anti-hexokinase (1:1000), mouse anti-α1 subunit of the Na+ /K+ ATPase (1:2000), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:200), and rabbit anti-α2 subunit of the Na+ /K+ ATPase (1:1000) were from Millipore (Temecula, CA).

    Article Title: Monomethyl Auristatin E Phosphate Inhibits Human Prostate Cancer Growth
    Article Snippet: Paragraph title: Western Blot Analysis ... Antibodies used include the following: mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and rabbit anti-cleaved poly ADP ribose polymerase (PARP) (Millipore, Billerica, MA).

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: Paragraph title: Western blotting and immunofluorescence ... Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences.

    Article Title: GABA transporters regulate tonic and synaptic GABAA receptor-mediated currents in the suprachiasmatic nucleus neurons
    Article Snippet: Paragraph title: Hypothalamic tissue collection and Western blot procedures. ... The loading control was mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) purified monoclonal antibody (MAB374; Millipore, Temecula, CA; 1:8,000).

    Protease Inhibitor:

    Article Title: Nitric Oxide Induces the Progression of Abdominal Aortic Aneurysms through the Matrix Metalloproteinase Inducer EMMPRIN
    Article Snippet: EDTA-free protease inhibitor mixture tablets were from Roche (Spain). .. Primary antibodies were obtained as follows: rabbit anti-MMP-13 from Calbiochem; goat-anti-human-MMP-13 and rabbit anti-inducible NO synthase (iNOS) from Santa Cruz Biotechnology (Santa Cruz, CA); rabbit anti-MMP-2, mouse anti-MMP-9, rabbit anti-MMP-12, and mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Chemicon (Millipore, Iberica, Spain); anti-FLAG, and goat-anti-EMMPRIN from Sigma-Aldrich; anti-ICAM-2, anti-von Willebrand factor, and anti-Mac-3 from Transduction Laboratories (BD Biosciences).

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: Cells were lysed either in M-PER Mammalian Protein Extraction Reagent (Thermo Scientific, Rockford, IL), supplemented with Halt protease inhibitor cocktail (Thermo Scientific), or in RIPA buffer (50 mM Tris, 150 mM NaCl, 0.1% SDS, 0.5% NaDeoxycholate, 1%NP40, 1 mM PMSF and Halt inhibitors added freshly) as indicated. .. Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences.

    Generated:

    Article Title: Monomethyl Auristatin E Phosphate Inhibits Human Prostate Cancer Growth
    Article Snippet: Results were generated using an Odyssey Infrared Imager (LI-COR Biosciences). .. Antibodies used include the following: mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and rabbit anti-cleaved poly ADP ribose polymerase (PARP) (Millipore, Billerica, MA).

    other:

    Article Title: Sorting nexin 27 regulates basal and stimulated brush border trafficking of NHE3
    Article Snippet: Mouse anti–glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and mouse anti-actin were from Sigma-Aldrich (St. Louis, MO).

    Imaging:

    Article Title: Glucose and glutamine handling in the Sertoli cells of transgenic rats overexpressing regucalcin: plasticity towards lactate production
    Article Snippet: A mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:10000, AB2302, Millipore, Darmstadt, Germany) antibody was used for protein loading control in all WB analyses. .. The membranes were incubated with ECL substrate (Bio-Rad) and immune-reactive proteins were scanned with the ChemiDoc™ MP Imaging System (Bio-Rad).

    Protein Concentration:

    Article Title: A20 Promotes Liver Regeneration by Decreasing SOCS3 Expression to Enhance IL-6/STAT3 Proliferative Signals
    Article Snippet: HepG2 and MPH whole cell lysates were recovered before and following IL-6, TNF and/or LPS treatment, and protein concentration determined ( ). .. Samples were analyzed by Western Blot (WB) using the following primary antibodies: rabbit anti-STAT3, rabbit anti-IκBα, mouse anti-βactin, (Santa Cruz Biotechnology, Inc., Santa Cruz, CA), rabbit anti-phospho-STAT3 (P-STAT3 Tyr705) (Cell Signaling Technology, Danvers, MA), chicken anti-TNFAIP3 (A20) (Abcam Inc., Cambridge, MA), mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Calbiochem/EMD Biosciences Inc., San Diego, CA), anti-hemagglutinin (HA) (Roche Applied Science, Indianapolis, IN); and secondary antibodies (Thermo Scientific, Rockford, IL).

    Article Title: GABA transporters regulate tonic and synaptic GABAA receptor-mediated currents in the suprachiasmatic nucleus neurons
    Article Snippet: Depending on the protein concentration in the lysate either 5 or 10 µg of total protein per 20-µl samples were loaded into the wells of a polyacrylamide gel. .. The loading control was mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) purified monoclonal antibody (MAB374; Millipore, Temecula, CA; 1:8,000).

    Affinity Purification:

    Article Title: Dynamic Regulation of Oct1 during Mitosis by Phosphorylation and Ubiquitination
    Article Snippet: The antibody was purified in two steps, first by blocking with unphosphorylated peptide to remove non-phospho-specific antibodies, then by affinity purification using the phosphorylated epitope. .. Mouse anti-pS10-H3 was obtained from Cell Signaling, and mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Chemicon.

    Article Title: Murine AKAP7 Has a 2′,5′-Phosphodiesterase Domain That Can Complement an Inactive Murine Coronavirus ns2 Gene
    Article Snippet: .. The following antibodies were used: mouse anti-ns2 monoclonal antibody (provided by Stuart Siddell, University of Bristol, United Kingdom), mouse monoclonal anti-Flag epitope (M2; Sigma); sheep anti-mouse IgG and horseradish peroxidase (HRP)-linked whole antibody (GE Healthcare); horse anti-mouse IgG HRP-linked antibody, goat anti-rabbit IgG HRP-linked antibody (Cell Signaling); Alexa Fluor 488 goat anti-mouse IgG, Alexa Fluor 594 goat anti-rabbit IgG secondary antibodies, and Alexa Fluor 488 goat anti-rabbit IgG (Invitrogen); mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (U.S. Biologicals); monoclonal anti-β-actin (A1978; Sigma-Aldrich); and custom-made affinity-purified rabbit polyclonal anti-AKAP7CD antibodies against peptide A (CQLLNEDEVNIGTDALLELK) and peptide B (KKQSNGYYHCESSIVIGEK) (Biosynthesis). ..

    Binding Assay:

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: .. Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences. .. For immunofluorescence, cells were grown on LabTek 8-well chamber slides, and fixed with 4% paraformaldehyde, followed by treatment with ice-cold methanol for 10 min. After permeabilization with 0.5% Triton X-100 in PBS for 3 min, cells were blocked with 10% normal goat serum and incubated with indicated primary antibodies.

    Immunofluorescence:

    Article Title: Vimentin Regulates Scribble Activity by Protecting It from Proteasomal Degradation
    Article Snippet: Monoclonal antibodies were mouse anti-GM130 clone 35 and anti-β-Catenin clone 14 (BD Biosciences Transduction Laboratories, Lexington, KY), mouse anti-vimentin clone V9 and anti-α-tubulin clone GTU-88 (Sigma-Aldrich), rat anti-HA clone 3F10 (Roche Diagnostics, Indianapolis, IN), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) clone 6C5 (Millipore Bioscience Research Reagents, Temecula, CA), mouse anti-keratin 18 clone C-04 (Abcam), and mouse anti-keratin 18 clone LDK18, a gift from Birgit E. Lane (Institute of Medical Biology, Immunos, Singapore). .. Control antibodies used in immunoprecipitations were normal mouse and goat immunoglobulin G (IgG) (Santa Cruz Biotechnology) Secondary antibodies used for immunofluorescence were donkey anti-mouse IgG Alexa Fluor 488 and 594, anti-goat IgG Alexa Fluor 488 and 594, and anti-rabbit IgG Alexa Fluor 488 (Invitrogen) and donkey anti-mouse IgG 7-amino-4-methylcoumarin-3-acetic acid (Jackson ImmunoResearch Laboratories, West Grove, PA).

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: Paragraph title: Western blotting and immunofluorescence ... Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences.

    Molecular Weight:

    Article Title: The glutamate transporter, GLAST, participates in a macromolecular complex that supports glutamate metabolism
    Article Snippet: Proteins (including rainbow molecular weight marker, Amersham) were resolved using 8% or 10% SDS-polyacrylamide gels and transferred to Immobilon FL polyvinylidene fluoride membranes (Millipore, Bedford, MA). .. Mouse anti-hexokinase (1:1000), mouse anti-α1 subunit of the Na+ /K+ ATPase (1:2000), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:200), and rabbit anti-α2 subunit of the Na+ /K+ ATPase (1:1000) were from Millipore (Temecula, CA).

    Fluorescence:

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences. .. Immunofluorescence was evaluated with a fluorescence microscope Axioplan2 (Zeiss, Sweden).

    Microscopy:

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences. .. Immunofluorescence was evaluated with a fluorescence microscope Axioplan2 (Zeiss, Sweden).

    Purification:

    Article Title: Dynamic Regulation of Oct1 during Mitosis by Phosphorylation and Ubiquitination
    Article Snippet: The antibody was purified in two steps, first by blocking with unphosphorylated peptide to remove non-phospho-specific antibodies, then by affinity purification using the phosphorylated epitope. .. Mouse anti-pS10-H3 was obtained from Cell Signaling, and mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Chemicon.

    Article Title: Localization and expression of GABA transporters in the suprachiasmatic nucleus
    Article Snippet: .. For a loading control a mouse anti-Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) purified monoclonal antibody (MAB374, Millipore, Temecula, CA, 1:8000) was applied followed by rabbit polyclonal anti-mouse IgG-HRP conjugated secondary antibody (AP160P, Millipore, 1:200,000). .. A molecular mass marker (kDa) was applied (# 161-0374, Bio-Rad, Life Science Research).

    Article Title: GABA transporters regulate tonic and synaptic GABAA receptor-mediated currents in the suprachiasmatic nucleus neurons
    Article Snippet: .. The loading control was mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) purified monoclonal antibody (MAB374; Millipore, Temecula, CA; 1:8,000). .. The bands were detected on the X-ray film following the incubation with a chemiluminescent substrate (catalog no. PI-34077; Thermo Scientific, Rockford, IL).

    Protein Extraction:

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: Nuclear extracts were prepared using NE-PER Nuclear Protein Extraction Kit from Thermo Scientific, to detect proteins presenting in the nuclei. .. Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences.

    Staining:

    Article Title: Septins Arrange F-Actin-Containing Fibers on the Chlamydia trachomatis Inclusion and Are Required for Normal Release of the Inclusion by Extrusion
    Article Snippet: The following primary antibodies were used in this study: mouse anti-chlamydia Hsp60 (c Hsp60; Enzo Life Sciences), mouse anti-α-tubulin (Sigma), rabbit antivimentin (Acris), mouse anti-phospho-Src Tyr 416 (pTyr-Src) (clone 9A6; Millipore) , rabbit anti-SEPT2 (11397-1-AP; Proteintech) or rabbit anti-SEPT2 (HPA018481; Sigma), rabbit anti-SEPT7 (H-120; Santa Cruz), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Millipore), mouse anti-β-actin (Sigma), rabbit anti-GFP (Cell Signaling), rabbit anti-SEPT9 (made by immunization with SEPT9 peptide), rabbit anti-SEPT11 (Abcam), and mouse anti-nonmuscle myosin IIB (clone 3H2; Abcam). .. Phalloidin Alexa Flour 546 (Life Technologies) was used to detect F actin, bisbenzimide Hoechst 33342 trihydrochloride (Sigma) was used to visualize host nuclei and bacterial DNA, and Sytox green nucleic acid stain (Life Technologies) was used to detect extruded chlamydial inclusions.

    SDS Page:

    Article Title: Glucose and glutamine handling in the Sertoli cells of transgenic rats overexpressing regucalcin: plasticity towards lactate production
    Article Snippet: Proteins (50 μg of each protein extract) were resolved in a 12% gel by SDS-PAGE and transferred to a PVDF membrane (Bio-Rad). .. A mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:10000, AB2302, Millipore, Darmstadt, Germany) antibody was used for protein loading control in all WB analyses.

    Article Title: Localization and expression of GABA transporters in the suprachiasmatic nucleus
    Article Snippet: The proteins were SDS-PAGE separated at 125V and electroblotted (40V for 1 hour) onto PVDF membranes. .. For a loading control a mouse anti-Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) purified monoclonal antibody (MAB374, Millipore, Temecula, CA, 1:8000) was applied followed by rabbit polyclonal anti-mouse IgG-HRP conjugated secondary antibody (AP160P, Millipore, 1:200,000).

    Article Title: GABA transporters regulate tonic and synaptic GABAA receptor-mediated currents in the suprachiasmatic nucleus neurons
    Article Snippet: The proteins were SDS-PAGE separated at 125 V and electroblotted (40 V for 1 h) onto polyvinylidene difluoride membranes. .. The loading control was mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) purified monoclonal antibody (MAB374; Millipore, Temecula, CA; 1:8,000).

    Software:

    Article Title: Glucose and glutamine handling in the Sertoli cells of transgenic rats overexpressing regucalcin: plasticity towards lactate production
    Article Snippet: A mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:10000, AB2302, Millipore, Darmstadt, Germany) antibody was used for protein loading control in all WB analyses. .. The density of the bands was obtained according to standard methods using the Image Lab 5.1 software (Bio‐Rad) and normalized by division with the respective GAPDH band density.

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: The optical density of each band was quantified using ImageQuant software. .. Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences.

    Marker:

    Article Title: Localization and expression of GABA transporters in the suprachiasmatic nucleus
    Article Snippet: For a loading control a mouse anti-Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) purified monoclonal antibody (MAB374, Millipore, Temecula, CA, 1:8000) was applied followed by rabbit polyclonal anti-mouse IgG-HRP conjugated secondary antibody (AP160P, Millipore, 1:200,000). .. A molecular mass marker (kDa) was applied (# 161-0374, Bio-Rad, Life Science Research).

    Article Title: The glutamate transporter, GLAST, participates in a macromolecular complex that supports glutamate metabolism
    Article Snippet: Proteins (including rainbow molecular weight marker, Amersham) were resolved using 8% or 10% SDS-polyacrylamide gels and transferred to Immobilon FL polyvinylidene fluoride membranes (Millipore, Bedford, MA). .. Mouse anti-hexokinase (1:1000), mouse anti-α1 subunit of the Na+ /K+ ATPase (1:2000), mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:200), and rabbit anti-α2 subunit of the Na+ /K+ ATPase (1:1000) were from Millipore (Temecula, CA).

    Lysis:

    Article Title: Protons Sensitize Epithelial Cells to Mesenchymal Transition
    Article Snippet: M-PER lysis buffer extracts only nuclear and cytoplasmic proteins. .. Mouse anti-vimentin and rabbit anti-fibronectin were purchased from Sigma-Aldrich; rabbit anti-phospho-Smad2 (S465/S467), rabbit anti-phospho-p53 and mouse anti-β-catenin were from Cell Signaling Technology Inc.; mouse anti-Smad7 from R & D mouse anti-TATA Box Binding Protein 1 (TBP1) and mouse anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Millipore; and mouse anti-E-cadherin from BD Biosciences.

    Fluorsave:

    Article Title: Nitric Oxide Induces the Progression of Abdominal Aortic Aneurysms through the Matrix Metalloproteinase Inducer EMMPRIN
    Article Snippet: Fluorsave coverslip mounting solution was from Calbiochem (CN Biosciences, UK). .. Primary antibodies were obtained as follows: rabbit anti-MMP-13 from Calbiochem; goat-anti-human-MMP-13 and rabbit anti-inducible NO synthase (iNOS) from Santa Cruz Biotechnology (Santa Cruz, CA); rabbit anti-MMP-2, mouse anti-MMP-9, rabbit anti-MMP-12, and mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Chemicon (Millipore, Iberica, Spain); anti-FLAG, and goat-anti-EMMPRIN from Sigma-Aldrich; anti-ICAM-2, anti-von Willebrand factor, and anti-Mac-3 from Transduction Laboratories (BD Biosciences).

    Article Title: c-Cbl and Cbl-b Act Redundantly to Protect Osteoclasts from Apoptosis and to Displace HDAC6 from ?-Tubulin, Stabilizing Microtubules and Podosomes
    Article Snippet: Mouse anti-β-tubulin, mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), mouse anti-HDAC6 and mouse anti-actin antibodies were obtained from Millipore Bioscience Research Reagents (Temecula, CA). .. HRP-conjugated anti-Myc and HRP-conjugated anti-hemagglutinin (HA) was purchased from Roche Applied Sciences (Indianapolis, IN). zVAD-FMK and FluorSave were purchased from Calbiochem (San Diego, CA).

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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
    Average 90 stars, based on 2 article reviews
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    mouse anti bax monoclonal - by Bioz Stars, 2020-03
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    93
    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: 93/100, 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, 2020-03
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    99
    Millipore anti glyceraldehyde 3 phosphate dehydrogenase
    Na v 1.1 levels are reduced in Scn1a RX/+ mice, and this reduction is not prevented by tau ablation. Levels of Na v 1.1 and total sodium channels (pan Na v ) in the parietal cortex of 8-month-old mice were determined by Western blot analysis. <t>Glyceraldehyde-3-phosphate</t> dehydrogenase (GAPDH) levels were used as a loading control. (A) Representative Western blot. (B) Quantification of Western blot signals (n = 5–7 mice per genotype). The average Na v 1.1 to pan Na v ratio in Scn1a +/+ / Tau +/+ mice was arbitrarily defined as 1.0. ***p
    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
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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

    Na v 1.1 levels are reduced in Scn1a RX/+ mice, and this reduction is not prevented by tau ablation. Levels of Na v 1.1 and total sodium channels (pan Na v ) in the parietal cortex of 8-month-old mice were determined by Western blot analysis. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) levels were used as a loading control. (A) Representative Western blot. (B) Quantification of Western blot signals (n = 5–7 mice per genotype). The average Na v 1.1 to pan Na v ratio in Scn1a +/+ / Tau +/+ mice was arbitrarily defined as 1.0. ***p

    Journal: Annals of Neurology

    Article Title: Tau Reduction Prevents Disease in a Mouse Model of Dravet Syndrome

    doi: 10.1002/ana.24230

    Figure Lengend Snippet: Na v 1.1 levels are reduced in Scn1a RX/+ mice, and this reduction is not prevented by tau ablation. Levels of Na v 1.1 and total sodium channels (pan Na v ) in the parietal cortex of 8-month-old mice were determined by Western blot analysis. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) levels were used as a loading control. (A) Representative Western blot. (B) Quantification of Western blot signals (n = 5–7 mice per genotype). The average Na v 1.1 to pan Na v ratio in Scn1a +/+ / Tau +/+ mice was arbitrarily defined as 1.0. ***p

    Article Snippet: After blocking for 1 hour in 5% bovine serum albumin diluted in Tris-buffered saline (BSA-TBS), membranes were incubated overnight at 4°C in anti-Nav 1.1 (1:1,000; Alomone Labs, Jerusalem, Israel), anti–pan-sodium channel (Pan Nav, 1:1,000; Sigma), anti–glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:10,000; Millipore, Billerica, MA), anti-tau clone Tau-5 (1:3,000; Life Technologies), anti-tau clone EP2456Y (1:1,000; Millipore), anti–phospho-tau Ser 396/404 clone PHF-1 (1:3,000, a gift from Dr P. Davies), anti–phospho-tau Thr231 clone CP9 (1:25, a gift from Dr P. Davies), or anti–phospho-PHF-tau pSer202+Thr205 clone AT8 (1:80; Thermo Scientific, Waltham, MA).

    Techniques: Mouse Assay, Western Blot

    Cortical levels of total and phosphorylated tau are not altered in Scn1a RX/+ mice. Levels of phospho-tau (PHF-1, Ser396/Ser404; AT8, Ser202/Thr205; CP9, Thr231) and total tau (Tau-5, EP2456Y) in the parietal cortex of 8-month-old mice of the indicated genotypes were determined by Western blot analysis. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. (A) Representative Western blot. (B) Quantification of Western blot signals (n = 5–7 mice per genotype) revealed no statistically significant differences between Scn1a RX/+ /Tau +/+ and Scn1a +/+ / Tau +/+ mice (Student t test). Average phospho-tau to EP2456Y ratios (PHF-1, AT8, CP9) or average total tau levels (Tau-5, EP2456Y) in Scn1a +/+ / Tau +/+ mice were arbitrarily defined as 1.0. Values represent mean ± standard error of the mean.

    Journal: Annals of Neurology

    Article Title: Tau Reduction Prevents Disease in a Mouse Model of Dravet Syndrome

    doi: 10.1002/ana.24230

    Figure Lengend Snippet: Cortical levels of total and phosphorylated tau are not altered in Scn1a RX/+ mice. Levels of phospho-tau (PHF-1, Ser396/Ser404; AT8, Ser202/Thr205; CP9, Thr231) and total tau (Tau-5, EP2456Y) in the parietal cortex of 8-month-old mice of the indicated genotypes were determined by Western blot analysis. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. (A) Representative Western blot. (B) Quantification of Western blot signals (n = 5–7 mice per genotype) revealed no statistically significant differences between Scn1a RX/+ /Tau +/+ and Scn1a +/+ / Tau +/+ mice (Student t test). Average phospho-tau to EP2456Y ratios (PHF-1, AT8, CP9) or average total tau levels (Tau-5, EP2456Y) in Scn1a +/+ / Tau +/+ mice were arbitrarily defined as 1.0. Values represent mean ± standard error of the mean.

    Article Snippet: After blocking for 1 hour in 5% bovine serum albumin diluted in Tris-buffered saline (BSA-TBS), membranes were incubated overnight at 4°C in anti-Nav 1.1 (1:1,000; Alomone Labs, Jerusalem, Israel), anti–pan-sodium channel (Pan Nav, 1:1,000; Sigma), anti–glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:10,000; Millipore, Billerica, MA), anti-tau clone Tau-5 (1:3,000; Life Technologies), anti-tau clone EP2456Y (1:1,000; Millipore), anti–phospho-tau Ser 396/404 clone PHF-1 (1:3,000, a gift from Dr P. Davies), anti–phospho-tau Thr231 clone CP9 (1:25, a gift from Dr P. Davies), or anti–phospho-PHF-tau pSer202+Thr205 clone AT8 (1:80; Thermo Scientific, Waltham, MA).

    Techniques: Mouse Assay, Western Blot