cleaved caspase3  (Cell Signaling Technology Inc)

 
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    Name:
    Cleaved Caspase 3 Asp175 D3E9 Rabbit mAb PE Conjugate
    Description:
    This Cell Signaling Technology antibody is conjugated to phycoerythrin PE and tested in house for direct flow cytometry analysis in human cells The antibody is expected to exhibit the same species cross reactivity as the unconjugated Cleaved Caspase 3 Asp175 D3E9 Rabbit mAb 9579
    Catalog Number:
    12768
    Price:
    None
    Applications:
    Flow Cytometry
    Category:
    Antibody Conjugates
    Source:
    Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp175 of human caspase-3 protein.
    Reactivity:
    Human
    Buy from Supplier


    Structured Review

    Cell Signaling Technology Inc cleaved caspase3
    RhoA activity affects cell survival. (A) K562 cells were transfected with MOCK, RhoAN19 or RhoAL63 vectors followed by treatment with 1µM imatinib for the indicated time points in hours. Cell lysates were assessed by western blot for the presence of <t>pro-caspase3</t> and cleaved active caspase3. Beta actin was used as a loading control. Corresponding densitometric analysis indicates the mean±s.e.m of three independent experiments, (p
    This Cell Signaling Technology antibody is conjugated to phycoerythrin PE and tested in house for direct flow cytometry analysis in human cells The antibody is expected to exhibit the same species cross reactivity as the unconjugated Cleaved Caspase 3 Asp175 D3E9 Rabbit mAb 9579
    https://www.bioz.com/result/cleaved caspase3/product/Cell Signaling Technology Inc
    Average 90 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    cleaved caspase3 - by Bioz Stars, 2020-02
    90/100 stars

    Images

    1) Product Images from "Increased Cytoplasmic Localization of p27kip1 and Its Modulation of RhoA Activity during Progression of Chronic Myeloid Leukemia"

    Article Title: Increased Cytoplasmic Localization of p27kip1 and Its Modulation of RhoA Activity during Progression of Chronic Myeloid Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0076527

    RhoA activity affects cell survival. (A) K562 cells were transfected with MOCK, RhoAN19 or RhoAL63 vectors followed by treatment with 1µM imatinib for the indicated time points in hours. Cell lysates were assessed by western blot for the presence of pro-caspase3 and cleaved active caspase3. Beta actin was used as a loading control. Corresponding densitometric analysis indicates the mean±s.e.m of three independent experiments, (p
    Figure Legend Snippet: RhoA activity affects cell survival. (A) K562 cells were transfected with MOCK, RhoAN19 or RhoAL63 vectors followed by treatment with 1µM imatinib for the indicated time points in hours. Cell lysates were assessed by western blot for the presence of pro-caspase3 and cleaved active caspase3. Beta actin was used as a loading control. Corresponding densitometric analysis indicates the mean±s.e.m of three independent experiments, (p

    Techniques Used: Activity Assay, Transfection, Western Blot

    2) Product Images from "LIM and SH3 protein 1 regulates cell growth and chemosensitivity of human glioblastoma via the PI3K/AKT pathway"

    Article Title: LIM and SH3 protein 1 regulates cell growth and chemosensitivity of human glioblastoma via the PI3K/AKT pathway

    Journal: BMC Cancer

    doi: 10.1186/s12885-018-4649-2

    LASP1 knockdown reduces tumor growth and promotes chemosensitivity in vivo. a U87 cells were transfected with lentivirus-based shRNA against LASP1, and western blotting was performed to determine knockdown efficiency; ( b ) qRT-PCR assay showed inhibition of LASP1 at the mRNA level; ( c ) stable construction tumor cells were injected subcutaneously into the back of nude mice to detect proliferative ability and chemosensitivity to TMZ. Tumor volume and weight were measured at 30 days after injection and expressed as the mean ± SD, n = 5; ( d ) proliferative ability of tumor cells was measured by Ki-67 and apoptosis rate was derived by cleaved caspase3 immunohistochemical staining, Scale bar, 50 μm. * P
    Figure Legend Snippet: LASP1 knockdown reduces tumor growth and promotes chemosensitivity in vivo. a U87 cells were transfected with lentivirus-based shRNA against LASP1, and western blotting was performed to determine knockdown efficiency; ( b ) qRT-PCR assay showed inhibition of LASP1 at the mRNA level; ( c ) stable construction tumor cells were injected subcutaneously into the back of nude mice to detect proliferative ability and chemosensitivity to TMZ. Tumor volume and weight were measured at 30 days after injection and expressed as the mean ± SD, n = 5; ( d ) proliferative ability of tumor cells was measured by Ki-67 and apoptosis rate was derived by cleaved caspase3 immunohistochemical staining, Scale bar, 50 μm. * P

    Techniques Used: In Vivo, Transfection, shRNA, Western Blot, Quantitative RT-PCR, Inhibition, Injection, Mouse Assay, Derivative Assay, Immunohistochemistry, Staining

    Related Articles

    Proton NMR:

    Article Title: Asiatic Acid Inhibits Pro-Angiogenic Effects of VEGF and Human Gliomas in Endothelial Cell Culture Models
    Article Snippet: Reagents Synthetic AsA, with structure confirmed by infrared and proton NMR spectra, was obtained from Sigma-Aldrich (St Louis, MO, USA). .. Primary antibodies for cleaved caspase 3, cleaved caspase 9, cleaved poly(ADP-ribose)polymerase (cPARP), pAkt-ser473, total Akt, Bad, and anti-rabbit peroxidase-conjugated secondary antibody were obtained from Cell Signaling (Beverly, MA, USA).

    Nucleic Acid Electrophoresis:

    Article Title: Protective Effects of Oleic Acid Against Palmitic Acid-Induced Apoptosis in Pancreatic AR42J Cells and Its Mechanisms
    Article Snippet: Western blot analysis Whole cell lysates were prepared by washing the cells twice with cold PBS followed by lysing the cells on ice with lysis buffer (150 mM NaCl, 50 mM Tris, 1 mM PMSF, 1 mM Na3 VO4 , 1% NP-40, 0.1% SDS, 0.5% deoxycholic acid, 1% protease inhibitor cocktail, pH 7.5) for 20 min. Proteins in cell lysates were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the proteins were transferred onto PVDF membrane. .. The membranes were incubated with 5% non-fat dry milk, and then incubated with antibodies towards Bcl-2, Bcl-xL, Mcl-1, Bak, cleaved caspase-3 (Asp175), and cleaved PARP (Asp214) for overnight at 4℃ Membranes were incubated with appropriate horseradish peroxidase-linked secondary antibody (Cell Signaling, Danvers, USA) at a dilution of 1:2,500.

    Article Title: Mechanism of the Susceptibility of Remodeled Pulmonary Vessels to Drug-Induced Cell Killing
    Article Snippet: Tissues were minced and homogenized, and protein gel electrophoresis samples were prepared as previously described. .. The membrane was blocked and incubated with antibodies for cleaved caspase‐3, LC3B, parkin (Cell Signaling Technology), Bcl‐xL , annexin A1, GATA4, and p62 (Syd Labs, Inc), and levels of proteins were detected using horseradish peroxidase‐linked secondary antibodies and an Enhanced Chemiluminescence System (Amersham Life Science).

    Article Title: Survivin Is a Therapeutic Target in Merkel Cell Carcinoma
    Article Snippet: .. Lysates were electrophoresed in 10% SDS–polyacrylamide gel electrophoresis, transferred to nitrocellulose membrane (Amersham), and reacted with specific antibodies CM2B4 (1:5000 dilution) , CM8E6 (1:500 dilution), cleaved PARP, cleaved caspase 3, survivin, XIAP, p53, BCL-2, BAX (1:1000 dilution, Cell Signaling Technology), E2F1, cyclin E (1:1000 dilution, Santa Cruz Biotechnology), LC3 (1:1000 dilution, Novus Biologicals), or α-tubulin (1:5000 dilution, Sigma) overnight at 4°C, followed by anti-mouse (1:5000 dilution, Amersham) or anti-rabbit immunoglobulin G (IgG)–horseradish peroxidase conjugates (1:3000 dilution, Cell Signaling Technology) for 1 hour at room temperature. .. Peroxidase activity was detected with Western Lightning Plus ECL reagent (PerkinElmer).

    BIA-KA:

    Article Title: LIM and SH3 protein 1 regulates cell growth and chemosensitivity of human glioblastoma via the PI3K/AKT pathway
    Article Snippet: Western blotting (WB) Cells were washed with 5% PBS 3 times and then lysed in RIPA lysis buffer for 1 h. Protein quantification was performed using BCA kits. .. The proteins were labeled with specific antibodies against LASP1 (1:1000; Chemicon, Temecula, CA, USA); cleaved poly (ADP-ribose) polymerase (PARP), cleaved caspase3, caspase3,p-PI3K, PI3K, p-Akt(Ser473), Akt (1:1000, Cell Signaling Technology, Danvers, MA, USA); and GAPDH (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA).

    Autoradiography:

    Article Title: LIM and SH3 protein 1 regulates cell growth and chemosensitivity of human glioblastoma via the PI3K/AKT pathway
    Article Snippet: The proteins were labeled with specific antibodies against LASP1 (1:1000; Chemicon, Temecula, CA, USA); cleaved poly (ADP-ribose) polymerase (PARP), cleaved caspase3, caspase3,p-PI3K, PI3K, p-Akt(Ser473), Akt (1:1000, Cell Signaling Technology, Danvers, MA, USA); and GAPDH (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA). .. Autoradiography signals were quantified and analyzed using Quantity One software (Bio-Rad, Hercules, CA, USA).

    Cytometry:

    Article Title: Platycodin D inhibits platelet function and thrombus formation through inducing internalization of platelet glycoprotein receptors
    Article Snippet: In brief, washed human platelets were treated with different doses of PD followed by measuring the surface expression of Annexin-V by flow cytometry using Annexin V Apoptosis Detection Kit (eBioscience, San Diego, CA, USA) or cleaved caspase-3 expression by western blot. .. Cleaved caspase-3 expression (rabbit monoclonal antibody against caspase-3, Cell Signaling Technology) was analyzed using Image J software and presented as a ratio relative to full length caspase-3.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Asiatic Acid Inhibits Pro-Angiogenic Effects of VEGF and Human Gliomas in Endothelial Cell Culture Models
    Article Snippet: Primary antibodies for cleaved caspase 3, cleaved caspase 9, cleaved poly(ADP-ribose)polymerase (cPARP), pAkt-ser473, total Akt, Bad, and anti-rabbit peroxidase-conjugated secondary antibody were obtained from Cell Signaling (Beverly, MA, USA). .. Human VEGF ELISA kit was from R & D systems (Minneapolis, MN, USA). α-tubulin antibody was from Neomarkers (Fremont, CA, USA).

    Incubation:

    Article Title: Increased Cytoplasmic Localization of p27kip1 and Its Modulation of RhoA Activity during Progression of Chronic Myeloid Leukemia
    Article Snippet: Following incubation with primary and HRP conjugated secondary antibodies, the blots were developed using Enhanced Chemiluminescent substrate (Pierce). .. Primary antibodies against phospho p44/42 MAPK, p44/42 MAPK, phospho SAPK/JNK, SAPK/JNK, phospho c-jun and c-jun and cleaved caspase3 were purchased from Cell Signaling Technology (Denvers, USA).

    Article Title: Protective Effects of Oleic Acid Against Palmitic Acid-Induced Apoptosis in Pancreatic AR42J Cells and Its Mechanisms
    Article Snippet: .. The membranes were incubated with 5% non-fat dry milk, and then incubated with antibodies towards Bcl-2, Bcl-xL, Mcl-1, Bak, cleaved caspase-3 (Asp175), and cleaved PARP (Asp214) for overnight at 4℃ Membranes were incubated with appropriate horseradish peroxidase-linked secondary antibody (Cell Signaling, Danvers, USA) at a dilution of 1:2,500. .. Immunoreactive bands were detected using the Enhanced chemiluminescence Western blotting detection system from GE Healthcare (Buckinghamshire, UK) according to manufacturer's instructions.

    Article Title: ROCK Inhibitor Y-27632 Suppresses Dissociation-Induced Apoptosis of Murine Prostate Stem/Progenitor Cells and Increases Their Cloning Efficiency
    Article Snippet: .. The membrane was blocked in 5% skim milk, and subsequently incubated with primary antibodies against β-actin (Sigma, St. Louis, MO), ROCK1 (Santa Cruz Biotechnology, Santa Cruz, CA), ROCK2 (BD Biosciences, Two Oak Park, Bedford, MA), MYPT, phospho-MYPT Thr853, Cleaved caspase 3, PTEN, phospho-PTEN Thr-380, AKT, phospho-AKT Thr473 (Cell Signaling Technology, Beverly, MA) at 4°C overnight followed by incubation with peroxidase-conjugated goat anti-mouse IgG or goat anti-rabbit IgG (Jackson ImmunoResearch, Inc., West Grove, PA), and developed with Pierce ECL reagent (Thermal Scientific, Rockford, IL). .. Annexin V staining Primary murine prostate cells were cultured in PrEC media in 6-cm Petri dishes with 10 µM Y-27632 for 6 days till cells were approximately 30–40% confluent.

    Article Title: Mechanism of the Susceptibility of Remodeled Pulmonary Vessels to Drug-Induced Cell Killing
    Article Snippet: .. The membrane was blocked and incubated with antibodies for cleaved caspase‐3, LC3B, parkin (Cell Signaling Technology), Bcl‐xL , annexin A1, GATA4, and p62 (Syd Labs, Inc), and levels of proteins were detected using horseradish peroxidase‐linked secondary antibodies and an Enhanced Chemiluminescence System (Amersham Life Science). .. Cell Culture Experiments Human PASMCs were purchased from ScienCell Research Laboratories and Cell Applications, Inc and were cultured in accordance with the manufacturers' instructions in 5% CO2 at 37°C.

    Article Title: A receptor tyrosine kinase ROR1 inhibitor (KAN0439834) induced significant apoptosis of pancreatic cells which was enhanced by erlotinib and ibrutinib
    Article Snippet: .. Filters were incubated with rabbit anti-PARP, cleaved caspase-3, MCL-1 and Bcl-xL antibodies (Cell Signaling Technology) overnight at 4°C, and subsequently with a secondary peroxidase-conjugated goat anti-rabbit antibody. ..

    Article Title: BET inhibitor OTX015 targets BRD2 and BRD4 and decreases c-MYC in acute leukemia cells
    Article Snippet: .. Cells were incubated with cleaved caspase-3 antibody (rabbit monoclonal Ab 9664; Cell Signaling Technology) or cytochrome c antibody (mouse monoclonal Ab 556432; Becton Dickinson) overnight at 4°C. .. Secondary antibodies were goat anti-rabbit and goat anti-mouse IgG coupled with Alexa 568 (red) or Alexa 488 (green) fluorochromes (Life Technologies) incubated at 37°C for 2h.

    Activity Assay:

    Article Title: Iron-Mediated Lysosomal Membrane Permeabilization in Ethanol-Induced Hepatic Oxidative Damage and Apoptosis: Protective Effects of Quercetin
    Article Snippet: Cathepsin B activity assay kit and apoptosis assay kit were from BioVision (USA) and KenGEN BioTECH (Nanjing, China), respectively. .. Cleaved caspase-3 antibody and cytochrome C antibody were purchased from CST (USA).

    Article Title: Survivin Is a Therapeutic Target in Merkel Cell Carcinoma
    Article Snippet: Lysates were electrophoresed in 10% SDS–polyacrylamide gel electrophoresis, transferred to nitrocellulose membrane (Amersham), and reacted with specific antibodies CM2B4 (1:5000 dilution) , CM8E6 (1:500 dilution), cleaved PARP, cleaved caspase 3, survivin, XIAP, p53, BCL-2, BAX (1:1000 dilution, Cell Signaling Technology), E2F1, cyclin E (1:1000 dilution, Santa Cruz Biotechnology), LC3 (1:1000 dilution, Novus Biologicals), or α-tubulin (1:5000 dilution, Sigma) overnight at 4°C, followed by anti-mouse (1:5000 dilution, Amersham) or anti-rabbit immunoglobulin G (IgG)–horseradish peroxidase conjugates (1:3000 dilution, Cell Signaling Technology) for 1 hour at room temperature. .. Peroxidase activity was detected with Western Lightning Plus ECL reagent (PerkinElmer).

    Expressing:

    Article Title: Protective Effects of Oleic Acid Against Palmitic Acid-Induced Apoptosis in Pancreatic AR42J Cells and Its Mechanisms
    Article Snippet: The membranes were incubated with 5% non-fat dry milk, and then incubated with antibodies towards Bcl-2, Bcl-xL, Mcl-1, Bak, cleaved caspase-3 (Asp175), and cleaved PARP (Asp214) for overnight at 4℃ Membranes were incubated with appropriate horseradish peroxidase-linked secondary antibody (Cell Signaling, Danvers, USA) at a dilution of 1:2,500. .. The expression level of each protein was normalized to that of a housekeeping protein, β-actin.

    Article Title: Platycodin D inhibits platelet function and thrombus formation through inducing internalization of platelet glycoprotein receptors
    Article Snippet: .. Cleaved caspase-3 expression (rabbit monoclonal antibody against caspase-3, Cell Signaling Technology) was analyzed using Image J software and presented as a ratio relative to full length caspase-3. ..

    Bradford Assay:

    Article Title: ROCK Inhibitor Y-27632 Suppresses Dissociation-Induced Apoptosis of Murine Prostate Stem/Progenitor Cells and Increases Their Cloning Efficiency
    Article Snippet: Protein concentrations were determined by Bradford Assay kit (BioRad, Hercules, CA). .. The membrane was blocked in 5% skim milk, and subsequently incubated with primary antibodies against β-actin (Sigma, St. Louis, MO), ROCK1 (Santa Cruz Biotechnology, Santa Cruz, CA), ROCK2 (BD Biosciences, Two Oak Park, Bedford, MA), MYPT, phospho-MYPT Thr853, Cleaved caspase 3, PTEN, phospho-PTEN Thr-380, AKT, phospho-AKT Thr473 (Cell Signaling Technology, Beverly, MA) at 4°C overnight followed by incubation with peroxidase-conjugated goat anti-mouse IgG or goat anti-rabbit IgG (Jackson ImmunoResearch, Inc., West Grove, PA), and developed with Pierce ECL reagent (Thermal Scientific, Rockford, IL).

    Western Blot:

    Article Title: Increased Cytoplasmic Localization of p27kip1 and Its Modulation of RhoA Activity during Progression of Chronic Myeloid Leukemia
    Article Snippet: Paragraph title: Western blot analysis ... Primary antibodies against phospho p44/42 MAPK, p44/42 MAPK, phospho SAPK/JNK, SAPK/JNK, phospho c-jun and c-jun and cleaved caspase3 were purchased from Cell Signaling Technology (Denvers, USA).

    Article Title: Protective Effects of Oleic Acid Against Palmitic Acid-Induced Apoptosis in Pancreatic AR42J Cells and Its Mechanisms
    Article Snippet: Paragraph title: Western blot analysis ... The membranes were incubated with 5% non-fat dry milk, and then incubated with antibodies towards Bcl-2, Bcl-xL, Mcl-1, Bak, cleaved caspase-3 (Asp175), and cleaved PARP (Asp214) for overnight at 4℃ Membranes were incubated with appropriate horseradish peroxidase-linked secondary antibody (Cell Signaling, Danvers, USA) at a dilution of 1:2,500.

    Article Title: ROCK Inhibitor Y-27632 Suppresses Dissociation-Induced Apoptosis of Murine Prostate Stem/Progenitor Cells and Increases Their Cloning Efficiency
    Article Snippet: Paragraph title: Western Blot analysis ... The membrane was blocked in 5% skim milk, and subsequently incubated with primary antibodies against β-actin (Sigma, St. Louis, MO), ROCK1 (Santa Cruz Biotechnology, Santa Cruz, CA), ROCK2 (BD Biosciences, Two Oak Park, Bedford, MA), MYPT, phospho-MYPT Thr853, Cleaved caspase 3, PTEN, phospho-PTEN Thr-380, AKT, phospho-AKT Thr473 (Cell Signaling Technology, Beverly, MA) at 4°C overnight followed by incubation with peroxidase-conjugated goat anti-mouse IgG or goat anti-rabbit IgG (Jackson ImmunoResearch, Inc., West Grove, PA), and developed with Pierce ECL reagent (Thermal Scientific, Rockford, IL).

    Article Title: Platycodin D inhibits platelet function and thrombus formation through inducing internalization of platelet glycoprotein receptors
    Article Snippet: In brief, washed human platelets were treated with different doses of PD followed by measuring the surface expression of Annexin-V by flow cytometry using Annexin V Apoptosis Detection Kit (eBioscience, San Diego, CA, USA) or cleaved caspase-3 expression by western blot. .. Cleaved caspase-3 expression (rabbit monoclonal antibody against caspase-3, Cell Signaling Technology) was analyzed using Image J software and presented as a ratio relative to full length caspase-3.

    Article Title: Mechanism of the Susceptibility of Remodeled Pulmonary Vessels to Drug-Induced Cell Killing
    Article Snippet: Paragraph title: Western Blot Analysis ... The membrane was blocked and incubated with antibodies for cleaved caspase‐3, LC3B, parkin (Cell Signaling Technology), Bcl‐xL , annexin A1, GATA4, and p62 (Syd Labs, Inc), and levels of proteins were detected using horseradish peroxidase‐linked secondary antibodies and an Enhanced Chemiluminescence System (Amersham Life Science).

    Article Title: GLP-1 analog liraglutide-induced cardiac dysfunction due to energetic starvation in heart failure with non-diabetic dilated cardiomyopathy
    Article Snippet: Paragraph title: Western blot analysis ... GLUT-4 antibody (#2213), cleaved caspase-3 antibody (#9661), phospho-AMPKα (Thr172) antibody (#2535), and LC3A/B (1/2) antibody (#12741), were purchased from Cell Signaling (Danvers, MA).

    Article Title: A receptor tyrosine kinase ROR1 inhibitor (KAN0439834) induced significant apoptosis of pancreatic cells which was enhanced by erlotinib and ibrutinib
    Article Snippet: Paragraph title: Western blot analysis ... Filters were incubated with rabbit anti-PARP, cleaved caspase-3, MCL-1 and Bcl-xL antibodies (Cell Signaling Technology) overnight at 4°C, and subsequently with a secondary peroxidase-conjugated goat anti-rabbit antibody.

    Article Title: LIM and SH3 protein 1 regulates cell growth and chemosensitivity of human glioblastoma via the PI3K/AKT pathway
    Article Snippet: Paragraph title: Western blotting (WB) ... The proteins were labeled with specific antibodies against LASP1 (1:1000; Chemicon, Temecula, CA, USA); cleaved poly (ADP-ribose) polymerase (PARP), cleaved caspase3, caspase3,p-PI3K, PI3K, p-Akt(Ser473), Akt (1:1000, Cell Signaling Technology, Danvers, MA, USA); and GAPDH (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA).

    Article Title: Survivin Is a Therapeutic Target in Merkel Cell Carcinoma
    Article Snippet: Lysates were electrophoresed in 10% SDS–polyacrylamide gel electrophoresis, transferred to nitrocellulose membrane (Amersham), and reacted with specific antibodies CM2B4 (1:5000 dilution) , CM8E6 (1:500 dilution), cleaved PARP, cleaved caspase 3, survivin, XIAP, p53, BCL-2, BAX (1:1000 dilution, Cell Signaling Technology), E2F1, cyclin E (1:1000 dilution, Santa Cruz Biotechnology), LC3 (1:1000 dilution, Novus Biologicals), or α-tubulin (1:5000 dilution, Sigma) overnight at 4°C, followed by anti-mouse (1:5000 dilution, Amersham) or anti-rabbit immunoglobulin G (IgG)–horseradish peroxidase conjugates (1:3000 dilution, Cell Signaling Technology) for 1 hour at room temperature. .. Peroxidase activity was detected with Western Lightning Plus ECL reagent (PerkinElmer).

    Flow Cytometry:

    Article Title: Platycodin D inhibits platelet function and thrombus formation through inducing internalization of platelet glycoprotein receptors
    Article Snippet: In brief, washed human platelets were treated with different doses of PD followed by measuring the surface expression of Annexin-V by flow cytometry using Annexin V Apoptosis Detection Kit (eBioscience, San Diego, CA, USA) or cleaved caspase-3 expression by western blot. .. Cleaved caspase-3 expression (rabbit monoclonal antibody against caspase-3, Cell Signaling Technology) was analyzed using Image J software and presented as a ratio relative to full length caspase-3.

    Protease Inhibitor:

    Article Title: Protective Effects of Oleic Acid Against Palmitic Acid-Induced Apoptosis in Pancreatic AR42J Cells and Its Mechanisms
    Article Snippet: Western blot analysis Whole cell lysates were prepared by washing the cells twice with cold PBS followed by lysing the cells on ice with lysis buffer (150 mM NaCl, 50 mM Tris, 1 mM PMSF, 1 mM Na3 VO4 , 1% NP-40, 0.1% SDS, 0.5% deoxycholic acid, 1% protease inhibitor cocktail, pH 7.5) for 20 min. Proteins in cell lysates were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the proteins were transferred onto PVDF membrane. .. The membranes were incubated with 5% non-fat dry milk, and then incubated with antibodies towards Bcl-2, Bcl-xL, Mcl-1, Bak, cleaved caspase-3 (Asp175), and cleaved PARP (Asp214) for overnight at 4℃ Membranes were incubated with appropriate horseradish peroxidase-linked secondary antibody (Cell Signaling, Danvers, USA) at a dilution of 1:2,500.

    Article Title: Survivin Is a Therapeutic Target in Merkel Cell Carcinoma
    Article Snippet: Cells were lysed in buffer [radioimmunoprecipitation assay or 10 mM tris-HCl (pH 8.0), 0.6% SDS] containing protease inhibitor cocktail (Roche). .. Lysates were electrophoresed in 10% SDS–polyacrylamide gel electrophoresis, transferred to nitrocellulose membrane (Amersham), and reacted with specific antibodies CM2B4 (1:5000 dilution) , CM8E6 (1:500 dilution), cleaved PARP, cleaved caspase 3, survivin, XIAP, p53, BCL-2, BAX (1:1000 dilution, Cell Signaling Technology), E2F1, cyclin E (1:1000 dilution, Santa Cruz Biotechnology), LC3 (1:1000 dilution, Novus Biologicals), or α-tubulin (1:5000 dilution, Sigma) overnight at 4°C, followed by anti-mouse (1:5000 dilution, Amersham) or anti-rabbit immunoglobulin G (IgG)–horseradish peroxidase conjugates (1:3000 dilution, Cell Signaling Technology) for 1 hour at room temperature.

    other:

    Article Title: The WT1/MVP-Mediated Stabilization on mTOR/AKT Axis Enhances the Effects of Cisplatin in Non-small Cell Lung Cancer by a Reformulated Yu Ping Feng San Herbal Preparation
    Article Snippet: The antibodies were obtained from the following sources: WT1 from Millipore (Billerica, MA, United States); MVP from Abcam (Cambridge, United Kingdom); mTOR, Rictor, p-AKT, cleaved-PARP, cleaved-caspase 3, horseradish peroxidase (HRP)-conjugated goat anti-rabbit antibody, HRP-conjugated goat anti-mouse antibody, and Alexa Fluor 555-conjugated goat anti-rabbit antibody from Cell Signaling Technology (Danvers, MA, United States).

    Imaging:

    Article Title: Survivin Is a Therapeutic Target in Merkel Cell Carcinoma
    Article Snippet: Lysates were electrophoresed in 10% SDS–polyacrylamide gel electrophoresis, transferred to nitrocellulose membrane (Amersham), and reacted with specific antibodies CM2B4 (1:5000 dilution) , CM8E6 (1:500 dilution), cleaved PARP, cleaved caspase 3, survivin, XIAP, p53, BCL-2, BAX (1:1000 dilution, Cell Signaling Technology), E2F1, cyclin E (1:1000 dilution, Santa Cruz Biotechnology), LC3 (1:1000 dilution, Novus Biologicals), or α-tubulin (1:5000 dilution, Sigma) overnight at 4°C, followed by anti-mouse (1:5000 dilution, Amersham) or anti-rabbit immunoglobulin G (IgG)–horseradish peroxidase conjugates (1:3000 dilution, Cell Signaling Technology) for 1 hour at room temperature. .. For quantitative immunoblotting, Odyssey Infrared Imaging System (LI-COR) was used with IRDye 800–conjugated secondary antibodies (1:5000 dilution, Rockland Immunochemicals).

    Recombinant:

    Article Title: Asiatic Acid Inhibits Pro-Angiogenic Effects of VEGF and Human Gliomas in Endothelial Cell Culture Models
    Article Snippet: Lyophilized recombinant human VEGF was purchased from Invitrogen/Gibco (Camarillo, CA, USA). .. Primary antibodies for cleaved caspase 3, cleaved caspase 9, cleaved poly(ADP-ribose)polymerase (cPARP), pAkt-ser473, total Akt, Bad, and anti-rabbit peroxidase-conjugated secondary antibody were obtained from Cell Signaling (Beverly, MA, USA).

    Article Title: A short-chain fatty acid, propionate, enhances the cytotoxic effect of cisplatin by modulating GPR41 signaling pathways in HepG2 cells
    Article Snippet: .. (S)-2-(4-chlorophenyl)-3,3-dimethyl-N-(5-phenylthiazol-2-yl) butanamide (CFMB) and TNF-α antagonist R-7050 [ ] (Calbiochem, Darmstadt, Germany); sodium propionate (NaP) (Sigma-Aldrich, St. Louis, MO, USA); pertussis toxin (PTX) (Wako, Osaka, Japan); gallein, cyclopropanecarboxylic acid (CPC) and trichostatin A (TSA) (Tokyo Chemical Industry, Tokyo, Japan); human recombinant TNF-α (PeproTech, Rocky Hill, NJ, USA); polyclonal rabbit antibodies against human β-actin (Abcam), acetyl-histone H3 (Lys9/Lys14), cleaved caspase-3 (Asp175), cleaved caspase-9 (all Cell Signaling Technology, Boston, MA USA), GPR41 (Abcam), and GPR43 (Bioss, Boston, MA, USA); monoclonal rabbit antibodies against HDAC1, HDAC4, HDAC5 (Cell Signaling Technology) and HDAC8 (Abcam); monoclonal mouse antibodies against HDAC3 and caspase-8 (Cell Signaling Technology); and horseradish peroxidase-conjugated anti-mouse, anti-goat and anti-rabbit immunoglobulins (Dako, Glostrup, Denmark) were used in the study. .. HepG2 cells, HuH-7 cells, JHH-4 cells and HLE cells, which are all human hepatoma cell lines, were obtained from the Japanese Collection of Research Bioresources Cell Bank.

    Immunofluorescence:

    Article Title: BET inhibitor OTX015 targets BRD2 and BRD4 and decreases c-MYC in acute leukemia cells
    Article Snippet: Paragraph title: Immunofluorescence ... Cells were incubated with cleaved caspase-3 antibody (rabbit monoclonal Ab 9664; Cell Signaling Technology) or cytochrome c antibody (mouse monoclonal Ab 556432; Becton Dickinson) overnight at 4°C.

    MTT Assay:

    Article Title: Capilliposide Isolated from Lysimachia capillipes Hemsl. Induces ROS Generation, Cell Cycle Arrest, and Apoptosis in Human Nonsmall Cell Lung Cancer Cell Lines
    Article Snippet: Dimethyl sulfoxide (DMSO), N-acetyl L-cysteine (NAC), cisplatin (DDP), 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphnyl-2H-tetrazolium bromide (MTT), phenylmethylsulfonyl fluoride (PMSF), 5-(and 6)-carboxy-2′7′-dichlorodihydrofluorescein diacetate (DCFDA) and the fluorescent dyes Hoechst 33342, and propidium iodide (PI) were all purchased from Sigma-Aldrich (St. Louis, MO, USA). .. The monoclonal antibodies against p53 (number 2527), Bax (#5023P), cleaved caspase-3 (#9661s), cleaved caspase-9 (#9505p), cytochrome C (#11940S), GAPDH (#2118), and horseradish peroxidase (HPR)-conjugated goat antirabbit secondary antibody (#7074P2) were obtained from Cell Signaling Technology (Cell Signaling Technology, MA, USA).

    Microscopy:

    Article Title: Mechanism of the Susceptibility of Remodeled Pulmonary Vessels to Drug-Induced Cell Killing
    Article Snippet: Western Blot Analysis Extrapulmonary arteries (left and right main branches) and intrapulmonary arteries (first‐order branch) were surgically dissected, and connective tissues were gently removed in ice‐cold PBS under a dissecting microscope. .. The membrane was blocked and incubated with antibodies for cleaved caspase‐3, LC3B, parkin (Cell Signaling Technology), Bcl‐xL , annexin A1, GATA4, and p62 (Syd Labs, Inc), and levels of proteins were detected using horseradish peroxidase‐linked secondary antibodies and an Enhanced Chemiluminescence System (Amersham Life Science).

    Article Title: BET inhibitor OTX015 targets BRD2 and BRD4 and decreases c-MYC in acute leukemia cells
    Article Snippet: Cells were incubated with cleaved caspase-3 antibody (rabbit monoclonal Ab 9664; Cell Signaling Technology) or cytochrome c antibody (mouse monoclonal Ab 556432; Becton Dickinson) overnight at 4°C. .. Images were acquired by immunofluorescence microscopy on a Zeiss Axiovert microscope with a Plan-Apochromat X63 N.A.1.4 oil immersion objective using the Axiovison software v4.2 (Carl Zeiss).

    Labeling:

    Article Title: LIM and SH3 protein 1 regulates cell growth and chemosensitivity of human glioblastoma via the PI3K/AKT pathway
    Article Snippet: .. The proteins were labeled with specific antibodies against LASP1 (1:1000; Chemicon, Temecula, CA, USA); cleaved poly (ADP-ribose) polymerase (PARP), cleaved caspase3, caspase3,p-PI3K, PI3K, p-Akt(Ser473), Akt (1:1000, Cell Signaling Technology, Danvers, MA, USA); and GAPDH (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA). .. Autoradiography signals were quantified and analyzed using Quantity One software (Bio-Rad, Hercules, CA, USA).

    SDS Page:

    Article Title: Increased Cytoplasmic Localization of p27kip1 and Its Modulation of RhoA Activity during Progression of Chronic Myeloid Leukemia
    Article Snippet: Equal amounts of protein was then run on parallel lanes of an SDS PAGE and blotted on PVDF membrane (GE Amersham, UK). .. Primary antibodies against phospho p44/42 MAPK, p44/42 MAPK, phospho SAPK/JNK, SAPK/JNK, phospho c-jun and c-jun and cleaved caspase3 were purchased from Cell Signaling Technology (Denvers, USA).

    Article Title: Protective Effects of Oleic Acid Against Palmitic Acid-Induced Apoptosis in Pancreatic AR42J Cells and Its Mechanisms
    Article Snippet: Western blot analysis Whole cell lysates were prepared by washing the cells twice with cold PBS followed by lysing the cells on ice with lysis buffer (150 mM NaCl, 50 mM Tris, 1 mM PMSF, 1 mM Na3 VO4 , 1% NP-40, 0.1% SDS, 0.5% deoxycholic acid, 1% protease inhibitor cocktail, pH 7.5) for 20 min. Proteins in cell lysates were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the proteins were transferred onto PVDF membrane. .. The membranes were incubated with 5% non-fat dry milk, and then incubated with antibodies towards Bcl-2, Bcl-xL, Mcl-1, Bak, cleaved caspase-3 (Asp175), and cleaved PARP (Asp214) for overnight at 4℃ Membranes were incubated with appropriate horseradish peroxidase-linked secondary antibody (Cell Signaling, Danvers, USA) at a dilution of 1:2,500.

    Article Title: ROCK Inhibitor Y-27632 Suppresses Dissociation-Induced Apoptosis of Murine Prostate Stem/Progenitor Cells and Increases Their Cloning Efficiency
    Article Snippet: Protein was separated by 8% or 12% SDS/PAGE and transferred onto a PVDF membrane (Amersham Biosciences, Arlington Heights, IL). .. The membrane was blocked in 5% skim milk, and subsequently incubated with primary antibodies against β-actin (Sigma, St. Louis, MO), ROCK1 (Santa Cruz Biotechnology, Santa Cruz, CA), ROCK2 (BD Biosciences, Two Oak Park, Bedford, MA), MYPT, phospho-MYPT Thr853, Cleaved caspase 3, PTEN, phospho-PTEN Thr-380, AKT, phospho-AKT Thr473 (Cell Signaling Technology, Beverly, MA) at 4°C overnight followed by incubation with peroxidase-conjugated goat anti-mouse IgG or goat anti-rabbit IgG (Jackson ImmunoResearch, Inc., West Grove, PA), and developed with Pierce ECL reagent (Thermal Scientific, Rockford, IL).

    Article Title: GLP-1 analog liraglutide-induced cardiac dysfunction due to energetic starvation in heart failure with non-diabetic dilated cardiomyopathy
    Article Snippet: Samples of heart tissue lysate were resolved on SDS-PAGE according to a standard protocol. .. GLUT-4 antibody (#2213), cleaved caspase-3 antibody (#9661), phospho-AMPKα (Thr172) antibody (#2535), and LC3A/B (1/2) antibody (#12741), were purchased from Cell Signaling (Danvers, MA).

    Article Title: LIM and SH3 protein 1 regulates cell growth and chemosensitivity of human glioblastoma via the PI3K/AKT pathway
    Article Snippet: Equal amounts of proteins were separated by SDS-PAGE and transferred onto polyvinylidene difluoride membranes (Amersham Pharmacia Biotech, Amersham, UK). .. The proteins were labeled with specific antibodies against LASP1 (1:1000; Chemicon, Temecula, CA, USA); cleaved poly (ADP-ribose) polymerase (PARP), cleaved caspase3, caspase3,p-PI3K, PI3K, p-Akt(Ser473), Akt (1:1000, Cell Signaling Technology, Danvers, MA, USA); and GAPDH (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA).

    Plasmid Preparation:

    Article Title: BET inhibitor OTX015 targets BRD2 and BRD4 and decreases c-MYC in acute leukemia cells
    Article Snippet: Cells were incubated with cleaved caspase-3 antibody (rabbit monoclonal Ab 9664; Cell Signaling Technology) or cytochrome c antibody (mouse monoclonal Ab 556432; Becton Dickinson) overnight at 4°C. .. Nuclei were counterstained with DAPI (Vector).

    Software:

    Article Title: Protective Effects of Oleic Acid Against Palmitic Acid-Induced Apoptosis in Pancreatic AR42J Cells and Its Mechanisms
    Article Snippet: The membranes were incubated with 5% non-fat dry milk, and then incubated with antibodies towards Bcl-2, Bcl-xL, Mcl-1, Bak, cleaved caspase-3 (Asp175), and cleaved PARP (Asp214) for overnight at 4℃ Membranes were incubated with appropriate horseradish peroxidase-linked secondary antibody (Cell Signaling, Danvers, USA) at a dilution of 1:2,500. .. The bands were quantified using ImageQuant software.

    Article Title: Platycodin D inhibits platelet function and thrombus formation through inducing internalization of platelet glycoprotein receptors
    Article Snippet: .. Cleaved caspase-3 expression (rabbit monoclonal antibody against caspase-3, Cell Signaling Technology) was analyzed using Image J software and presented as a ratio relative to full length caspase-3. ..

    Article Title: GLP-1 analog liraglutide-induced cardiac dysfunction due to energetic starvation in heart failure with non-diabetic dilated cardiomyopathy
    Article Snippet: Bands were revealed using ECL select western blotting detection reagents (GE Healthcare, Buckinghamshire, UK), and band density was quantified using the Image J software. .. GLUT-4 antibody (#2213), cleaved caspase-3 antibody (#9661), phospho-AMPKα (Thr172) antibody (#2535), and LC3A/B (1/2) antibody (#12741), were purchased from Cell Signaling (Danvers, MA).

    Article Title: A receptor tyrosine kinase ROR1 inhibitor (KAN0439834) induced significant apoptosis of pancreatic cells which was enhanced by erlotinib and ibrutinib
    Article Snippet: Filters were incubated with rabbit anti-PARP, cleaved caspase-3, MCL-1 and Bcl-xL antibodies (Cell Signaling Technology) overnight at 4°C, and subsequently with a secondary peroxidase-conjugated goat anti-rabbit antibody. .. Densitometric quantification was carried out using the ImageJ software (National Institute of Health, USA).

    Article Title: BET inhibitor OTX015 targets BRD2 and BRD4 and decreases c-MYC in acute leukemia cells
    Article Snippet: Cells were incubated with cleaved caspase-3 antibody (rabbit monoclonal Ab 9664; Cell Signaling Technology) or cytochrome c antibody (mouse monoclonal Ab 556432; Becton Dickinson) overnight at 4°C. .. Images were acquired by immunofluorescence microscopy on a Zeiss Axiovert microscope with a Plan-Apochromat X63 N.A.1.4 oil immersion objective using the Axiovison software v4.2 (Carl Zeiss).

    Article Title: LIM and SH3 protein 1 regulates cell growth and chemosensitivity of human glioblastoma via the PI3K/AKT pathway
    Article Snippet: The proteins were labeled with specific antibodies against LASP1 (1:1000; Chemicon, Temecula, CA, USA); cleaved poly (ADP-ribose) polymerase (PARP), cleaved caspase3, caspase3,p-PI3K, PI3K, p-Akt(Ser473), Akt (1:1000, Cell Signaling Technology, Danvers, MA, USA); and GAPDH (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA). .. Autoradiography signals were quantified and analyzed using Quantity One software (Bio-Rad, Hercules, CA, USA).

    In Situ:

    Article Title: Iron-Mediated Lysosomal Membrane Permeabilization in Ethanol-Induced Hepatic Oxidative Damage and Apoptosis: Protective Effects of Quercetin
    Article Snippet: In situ cell death detection kit was provided by Roche (Switzerland). .. Cleaved caspase-3 antibody and cytochrome C antibody were purchased from CST (USA).

    Radio Immunoprecipitation:

    Article Title: Survivin Is a Therapeutic Target in Merkel Cell Carcinoma
    Article Snippet: Cells were lysed in buffer [radioimmunoprecipitation assay or 10 mM tris-HCl (pH 8.0), 0.6% SDS] containing protease inhibitor cocktail (Roche). .. Lysates were electrophoresed in 10% SDS–polyacrylamide gel electrophoresis, transferred to nitrocellulose membrane (Amersham), and reacted with specific antibodies CM2B4 (1:5000 dilution) , CM8E6 (1:500 dilution), cleaved PARP, cleaved caspase 3, survivin, XIAP, p53, BCL-2, BAX (1:1000 dilution, Cell Signaling Technology), E2F1, cyclin E (1:1000 dilution, Santa Cruz Biotechnology), LC3 (1:1000 dilution, Novus Biologicals), or α-tubulin (1:5000 dilution, Sigma) overnight at 4°C, followed by anti-mouse (1:5000 dilution, Amersham) or anti-rabbit immunoglobulin G (IgG)–horseradish peroxidase conjugates (1:3000 dilution, Cell Signaling Technology) for 1 hour at room temperature.

    Apoptosis Assay:

    Article Title: Iron-Mediated Lysosomal Membrane Permeabilization in Ethanol-Induced Hepatic Oxidative Damage and Apoptosis: Protective Effects of Quercetin
    Article Snippet: Cathepsin B activity assay kit and apoptosis assay kit were from BioVision (USA) and KenGEN BioTECH (Nanjing, China), respectively. .. Cleaved caspase-3 antibody and cytochrome C antibody were purchased from CST (USA).

    Lysis:

    Article Title: Protective Effects of Oleic Acid Against Palmitic Acid-Induced Apoptosis in Pancreatic AR42J Cells and Its Mechanisms
    Article Snippet: Western blot analysis Whole cell lysates were prepared by washing the cells twice with cold PBS followed by lysing the cells on ice with lysis buffer (150 mM NaCl, 50 mM Tris, 1 mM PMSF, 1 mM Na3 VO4 , 1% NP-40, 0.1% SDS, 0.5% deoxycholic acid, 1% protease inhibitor cocktail, pH 7.5) for 20 min. Proteins in cell lysates were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the proteins were transferred onto PVDF membrane. .. The membranes were incubated with 5% non-fat dry milk, and then incubated with antibodies towards Bcl-2, Bcl-xL, Mcl-1, Bak, cleaved caspase-3 (Asp175), and cleaved PARP (Asp214) for overnight at 4℃ Membranes were incubated with appropriate horseradish peroxidase-linked secondary antibody (Cell Signaling, Danvers, USA) at a dilution of 1:2,500.

    Article Title: LIM and SH3 protein 1 regulates cell growth and chemosensitivity of human glioblastoma via the PI3K/AKT pathway
    Article Snippet: Western blotting (WB) Cells were washed with 5% PBS 3 times and then lysed in RIPA lysis buffer for 1 h. Protein quantification was performed using BCA kits. .. The proteins were labeled with specific antibodies against LASP1 (1:1000; Chemicon, Temecula, CA, USA); cleaved poly (ADP-ribose) polymerase (PARP), cleaved caspase3, caspase3,p-PI3K, PI3K, p-Akt(Ser473), Akt (1:1000, Cell Signaling Technology, Danvers, MA, USA); and GAPDH (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA).

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    Cell Signaling Technology Inc cleaved caspase 3
    DEX improved the apoptosis of rat renal tubular cells induced by acute stress. (a) The apoptosis of renal tissue was measured by the TUNEL Apoptosis kit, in which green cells represent TUNEL-positive apoptotic cells. TUNEL (green) detects apoptosis, DAPI (blue) locates the nucleus, and MERGE is the combination of TUNEL and DAPI. All fluorescence images were observed with a fluorescence microscope at a magnification of 200. Quantitative results of TUNEL analysis. Immunohistochemistry of cleaved <t>caspase</t> 3 protein: (b) C group, (c) AS group, and (d) D + AS group; bars = 50 μ m. Data are expressed as mean ± SEM ( n = 6). ∗∗ p
    Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 298 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    DEX improved the apoptosis of rat renal tubular cells induced by acute stress. (a) The apoptosis of renal tissue was measured by the TUNEL Apoptosis kit, in which green cells represent TUNEL-positive apoptotic cells. TUNEL (green) detects apoptosis, DAPI (blue) locates the nucleus, and MERGE is the combination of TUNEL and DAPI. All fluorescence images were observed with a fluorescence microscope at a magnification of 200. Quantitative results of TUNEL analysis. Immunohistochemistry of cleaved caspase 3 protein: (b) C group, (c) AS group, and (d) D + AS group; bars = 50 μ m. Data are expressed as mean ± SEM ( n = 6). ∗∗ p

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Dexmedetomidine Ameliorates Acute Stress-Induced Kidney Injury by Attenuating Oxidative Stress and Apoptosis through Inhibition of the ROS/JNK Signaling Pathway

    doi: 10.1155/2018/4035310

    Figure Lengend Snippet: DEX improved the apoptosis of rat renal tubular cells induced by acute stress. (a) The apoptosis of renal tissue was measured by the TUNEL Apoptosis kit, in which green cells represent TUNEL-positive apoptotic cells. TUNEL (green) detects apoptosis, DAPI (blue) locates the nucleus, and MERGE is the combination of TUNEL and DAPI. All fluorescence images were observed with a fluorescence microscope at a magnification of 200. Quantitative results of TUNEL analysis. Immunohistochemistry of cleaved caspase 3 protein: (b) C group, (c) AS group, and (d) D + AS group; bars = 50 μ m. Data are expressed as mean ± SEM ( n = 6). ∗∗ p

    Article Snippet: Therefore, the site of kidney injury was determined by measuring localization of cleaved caspase 3 in kidney tissue.

    Techniques: TUNEL Assay, Fluorescence, Microscopy, Immunohistochemistry

    ( A, B ) Western blot analysis was performed to detect the effect of melatonin on factors related to cell apoptosis. Melatonin administration upregulated the level of Bcl-2 but reduced the levels of Bax and cleavage caspase-3 following the onset of SAH (P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: Melatonin Upregulates Nuclear Factor Erythroid-2 Related Factor 2 (Nrf2) and Mediates Mitophagy to Protect Against Early Brain Injury After Subarachnoid Hemorrhage

    doi: 10.12659/MSM.909221

    Figure Lengend Snippet: ( A, B ) Western blot analysis was performed to detect the effect of melatonin on factors related to cell apoptosis. Melatonin administration upregulated the level of Bcl-2 but reduced the levels of Bax and cleavage caspase-3 following the onset of SAH (P

    Article Snippet: In addition, high protein levels of Bax and cleaved caspase-3 were observed in the SAH group, while the melatonin administration decreased the protein levels of Bax and cleaved caspase-3, indicating that melatonin prevented the apoptosis of neurons following the onset of SAH.

    Techniques: Western Blot

    Correlation of the serum levels of 24S-HC with the expression of cleaved caspase-3 at 6hr and 24hr after neonatal HI. A. Protein expression of cleaved caspase-3 in the cortices was measured by western blotting at the indicated time points and presented as the OD ratio to β-actin and normalized to an internal control (IC). Sham vs. HI, *p=0.0112 at 6hr; *p=0.0027 at 24hr; *p=0.0228 at 72hr (n=5–6 for sham animals, n=7–10 for HI animals from 6–72hr). B. Correlation of the serum levels of 24S-HC with the expression of cleaved caspase-3 at 6hr (top) and at 24hr (bottom). Sample number, R 2 and p values are shown in the graphs.

    Journal: Pediatric research

    Article Title: Up-regulation of cholesterol 24-hydroxylase following hypoxia-ischemia in neonatal mouse brain

    doi: 10.1038/pr.2018.49

    Figure Lengend Snippet: Correlation of the serum levels of 24S-HC with the expression of cleaved caspase-3 at 6hr and 24hr after neonatal HI. A. Protein expression of cleaved caspase-3 in the cortices was measured by western blotting at the indicated time points and presented as the OD ratio to β-actin and normalized to an internal control (IC). Sham vs. HI, *p=0.0112 at 6hr; *p=0.0027 at 24hr; *p=0.0228 at 72hr (n=5–6 for sham animals, n=7–10 for HI animals from 6–72hr). B. Correlation of the serum levels of 24S-HC with the expression of cleaved caspase-3 at 6hr (top) and at 24hr (bottom). Sample number, R 2 and p values are shown in the graphs.

    Article Snippet: It is interesting that the expression of CYP46A1 in the ipsilateral cortex was largely localized to the cells undergoing apoptotic cell death as visualized by the expression of cleaved caspase-3 ( ).

    Techniques: Expressing, Western Blot

    Up-regulation of CYP46A1 with a concomitant increase of 24S-HC in the ipsilateral cortex after neonatal HI. A. Protein expression of CYP46A1 was measured by western blotting at the indicated time points and presented as the OD ratio to β-actin and normalized to the values of sham 0hr (graph on the right, sham vs. HI: * p= 0.0339 at 6hr; * p=0.0026 at 24hr; n=5–6 for sham animals, n=6–12 for HI animals at 6–72hr). B. Immunofluorescent staining with anti-CYP46A1 antibody at 24hr after HI showed enhanced expression in the ipsilateral cortex than that in the contralateral side and in the sham animals (left panel s). CYP46A1 staining in the ipsi-cortex was largely co-localized with the expression of cleaved caspase-3 (middle panels). C. Increased production of 24S-HC (ng/mg tissue wet weight) in the ipsilateral cortex at 6hr and 24hr after HI (sham vs. HI: *p= 0.0167 at 6hr; *p=0.0085 at 24hr; n=3–6 for sham animals, n=5–7 for HI animals at 6–72hr). The time course of the changes in 24S-HC in the sham-, contra- and ipsilateral cortex is shown on the right. At 6hr, 24S-HC in the ipsilateral cortex was higher than that in the contralateral side (ipsi- vs. contra-: *p=0.0493). #: Significant difference between the HI ipsi- and the sham animals only, but not between the HI ipsi- and HI contralateral hemisphere.

    Journal: Pediatric research

    Article Title: Up-regulation of cholesterol 24-hydroxylase following hypoxia-ischemia in neonatal mouse brain

    doi: 10.1038/pr.2018.49

    Figure Lengend Snippet: Up-regulation of CYP46A1 with a concomitant increase of 24S-HC in the ipsilateral cortex after neonatal HI. A. Protein expression of CYP46A1 was measured by western blotting at the indicated time points and presented as the OD ratio to β-actin and normalized to the values of sham 0hr (graph on the right, sham vs. HI: * p= 0.0339 at 6hr; * p=0.0026 at 24hr; n=5–6 for sham animals, n=6–12 for HI animals at 6–72hr). B. Immunofluorescent staining with anti-CYP46A1 antibody at 24hr after HI showed enhanced expression in the ipsilateral cortex than that in the contralateral side and in the sham animals (left panel s). CYP46A1 staining in the ipsi-cortex was largely co-localized with the expression of cleaved caspase-3 (middle panels). C. Increased production of 24S-HC (ng/mg tissue wet weight) in the ipsilateral cortex at 6hr and 24hr after HI (sham vs. HI: *p= 0.0167 at 6hr; *p=0.0085 at 24hr; n=3–6 for sham animals, n=5–7 for HI animals at 6–72hr). The time course of the changes in 24S-HC in the sham-, contra- and ipsilateral cortex is shown on the right. At 6hr, 24S-HC in the ipsilateral cortex was higher than that in the contralateral side (ipsi- vs. contra-: *p=0.0493). #: Significant difference between the HI ipsi- and the sham animals only, but not between the HI ipsi- and HI contralateral hemisphere.

    Article Snippet: It is interesting that the expression of CYP46A1 in the ipsilateral cortex was largely localized to the cells undergoing apoptotic cell death as visualized by the expression of cleaved caspase-3 ( ).

    Techniques: Expressing, Western Blot, Staining