Atorvastatin calcium, Mevalonic acid, GGPP, FPP, Propidium iodide (PI) and 2′,7′-dichlorofluorescein diacetate (DCFH-DA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was from Amresco (Solon, OH, USA). 5,5′,6,6′-tetrachloro-1, 1′,3,3′-tetraethylbenzimidazole-carbocyanide iodine (JC-1) and cell mitochondria isolation kit were obtained from Beyotime Biotechnology (Shanghai, China). Annexin V-FITC Apoptosis Detection Kit, as well as antibodies against p-pRb (S780) (1:1,000), p27 (1:1,000) and cyclinD1 (1:1,000) were from BD Biosciences Pharmingen (San Jose, CA, USA). Antibodies against pRb (1:1,000), cyclinB1 (1:2,000), cdc2 (1:1,000), caspase-3 (1:1,000), caspase-9 (1:1,000), poly (ADP-ribose) polymerase (PARP) (1:1,000), Cytochrome c (1:1,000), YAP (1:1,000), p-YAP (Ser127) (1:1,000), Rho A (1:1,000), β-actin (1:1,000), anti-mouse (1:2,000), anti-rabbit HRP-conjugated and anti-rabbit Alexa Fluor 488-conjugated secondary antibodies (1:2,000) were from Cell Signaling Technology (Danvers, MA, USA). Antibodies against Bcl-2 (1:1,000), Bax (1:1,000) and Lamin B (1:500) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Poly adp ribose polymerase parp
Manufactured by Cell Signaling Technology
Sourced in United States, United Kingdom
Poly (ADP-ribose) polymerase (PARP) is an enzyme that catalyzes the post-translational modification of proteins by adding long chains of ADP-ribose units. PARP plays a critical role in various cellular processes, including DNA repair, genomic stability, and cell death signaling.
Automatically generated - may contain errors
Lab products found in correlation
Caspase 3, by Cell Signaling Technology (52 mentions)
Cleaved caspase 3, by Cell Signaling Technology (34 mentions)
β actin, by Cell Signaling Technology (24 mentions)
Caspase 9, by Cell Signaling Technology (21 mentions)
Bcl 2, by Cell Signaling Technology (17 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (15 mentions)
Bcl xl, by Cell Signaling Technology (14 mentions)
Cleaved parp, by Cell Signaling Technology (12 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (11 mentions)
Caspase 8, by Cell Signaling Technology (9 mentions)
β actin, by Merck Group (9 mentions)
Gapdh, by Cell Signaling Technology (8 mentions)
Survivin, by Cell Signaling Technology (8 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (8 mentions)
P akt, by Cell Signaling Technology (7 mentions)
β actin, by Santa Cruz Biotechnology (7 mentions)
Propidium iodide, by Merck Group (7 mentions)
Cleaved caspase 9, by Cell Signaling Technology (6 mentions)
Cytochrome c, by Cell Signaling Technology (5 mentions)
Cyclin d1, by Abcam (5 mentions)
152 protocols using poly adp ribose polymerase parp
1
Atorvastatin Inhibits Proliferation and Induces Apoptosis in Cancer Cells
2
Protein Expression Analysis of Key Cellular Markers
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Cells were washed twice with ice-cold PBS and lysed in a radioimmunoprecipitation assay buffer [100 mM Tris-HCl (pH 8.0), 150 mM NaCl, 0.1% SDS, and 1% Triton X-100] at 4°C. The proteins in the resulting lysate were separated using sodium dodecyl sulphate polyacrylamide gel electrophoresis, and the resolved proteins were immunoblotted with antibodies against β-actin, nuclear factor erythroid 2-related factor 2 (Nrf2), B-cell lymphoma 2 (BCL2) and adenovirus E1B 19-kDa-interacting protein 3 (BNIP-3), p53, Slug (Santa Cruz Biotechnology, Santa Cruz, CA, USA), HIF-1α, α-Smooth muscle actin (α-SMA), transforming growth factor-β (TGF-β), N-cadherin, E-cadherin, Snail, vimentin, poly (ADP-ribose) polymerase (PARP; Cell Signaling Technology, Danvers, MA, USA), γH2A.X, cyclin D1, collagen-I (Abcam, Cambridge, UK), heme-oxidase 1 (HO-1; Enzo Life Sciences, Farmingdale, NY, USA), and differentiated embryonic chondrocyte gene 1 (DEC-1; Bethyl Laboratory, TX, USA).
3
Apoptosis Pathway Activation Protocol
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Caspase-9, caspase-8, caspase-3, cleaved-caspase-3, poly(ADP-ribose) polymerase (PARP), XIAP, cIAP-1, cIAP2, Bcl-2, Bcl-xL, and Bax were procured from Cell Signaling Technologies (Beverly, MA, USA) and the GAPDH antibody was procured from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, United States). Annexin V-FITC, propidium iodide staining solution, Hoechst33342Solution, BD Cytofix/Cytoperm Plus fixation and permeabilization solution kit(BD (Pharmingen San Jose, CA, USA). The Cell Counting Kit-8 (CCK-8) kit and N-acetyl cysteine (NAC) was obtained from Sigma-Aldrich (St. Louis, MO, United States). z-VAD-FMK was bought from Calbiochem (San Diego, CA, USA). CellROXGreen, MitoSOXRed, andThiolTracker Violet were purchased from Invitrogen (Waltham, MA, USA). Mitopotential kit was purchased from the EMD Millipore Corporation (Danvers, MA, USA).
4
Western Blot Analysis of Apoptosis Markers
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Following cell lysis by ice-cold radioimmunoprecipitation assay buffer, protein concentrations were determined. Equal quantities (30 µg) of protein were separated using 10% SDS-polyacrylamide gels, transferred to a polyvinylidene difluoride (PVDF) membrane and blocked with 5% bovine serum albumin (BSA). PVDF membranes were incubated overnight at 4°C with the following antibodies: CIP2A (dilution 1:500; cat. no. sc-80662; Santa Cruz Biotechnology, Inc., Dallas, CA, USA), AKT (dilution 1:1,000; cat. no. 9272S; Cell Signaling Technology, Inc., Danvers, MA, USA), phosphorylated (p)-AKT (dilution 1:1,000; cat. no. 4060S; Cell Signaling Technology), B cell lymphoma-2 (Bcl-2; dilution 1:1,000; cat. no. sc-492; Santa Cruz Biotechnology, Inc.), caspase-3 (dilution 1:1,000; cat. no. 9662S; Cell Signaling Technology), poly(ADP-Ribose) polymerase (PARP; dilution 1:1,000; cat. no. 5625T; Cell Signaling Technology), γ-H2AX (dilution 1:1,000; cat. no. ab2893; Abcam, Cambridge, UK) and β-actin (dilution 1:1,000; cat. no. sc-47778; Santa Cruz Biotechnology, Inc.). After being washed, the membranes were then incubated with horseradish peroxidase-conjugated secondary antibodies (anti-mouse IgG, dilution 1:1,000; cat. no. 7076S; Cell Signaling Technology; anti-rabbit IgG, dilution 1:1,000; cat. no. 7074S; Cell Signaling Technology) followed by enhanced chemiluminescence detection.
5
Cucurbitacin I Inhibits VEGF-Induced Angiogenesis
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JSI-124 (Cucurbitacin I) was purchased from Sigma. A 1 mg/ml JSI-124 stock solution was prepared in dimethyl sulfoxide (DMSO; Sigma), stored at −20°C and then diluted as needed in cell culture medium. Recombinant human VEGF165 was purchased from R&D Systems. Matrigel and transwell chambers were obtained from BD Biosciences. Antibodies against JAK2, STAT3, phospho-STAT3 (Ser727),VEGFR2, phospho-VEGFR2 (Tyr1175), Bcl-2, Bcl-xL, Caspase-3, GAPDH and poly (ADP-ribose) polymerase (PARP) were obtained from Cell Signaling Technology. Phospho-JAK2 (Y1007/Y1008) was purchased from Abcam.
6
Antibody-based Apoptosis Analysis
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Antibodies against caspase-3, -8, -9, and poly (ADP-ribose) polymerase (PARP) were obtained from Cell Signaling Technology (Beverly, MA, USA). Antibody against β-actin was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Fetal bovine serum (FBS) was from GIBCO (Gaithersburg, MD, USA). Caspase inhibitors were from Calciochem (San Diego, CA, USA). All other chemicals were obtained from Wako Pure Chemical Industries (Osaka, Japan).
7
Western Blot Analysis of Protein Expression
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Cells were lysed in lysis buffer (1.5% SDS, 1% NP-40, 10 mM Tris, pH 8.0, and 1 mM EDTA) containing a protease inhibitor cocktail (cOmplete™, Roche). Sonication was performed and the sample was then boiled at 95°C for 6 min. Total protein concentrations were measured using BCA assay (Sigma–Aldrich). Protein samples were separated by SDS/PAGE and transferred on to PVDF membranes. After blocking with 5% skim milk in TBS containing 0.1% Tween 20, membranes were incubated with antibodies specific for desmoglein1 (Dsg1), desmoglein3 (Dsg3), tubulin, actin (Santa Cruz Biotechnology, Santa Cruz, CA), poly(ADP-ribose) polymerase (PARP) (Cell Signaling Technology, Danvers, MA, U.S.A.), ubiquitinylated conjugates (FK2 clone, Enzo Life Sciences, lnc., Farmingdale, NY, U.S.A.), GAPDH, or Dsp (Thermo Fisher Scientific). Membranes were washed using TBS with 0.1% Tween-20 and incubated with secondary antibodies. Signals were detected using an ECL solution (Supersignal West Pico, Thermo Fisher Scientific).
8
Apoptosis Induction Protocol
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RPMI 1640, penicillin, streptomycin and PBS were purchased from Gibco, Lifetechnologies (Darmstadt, Germany). FCS was purchased from Biochrom (Berlin, Germany). RIPA buffer, protein inhibitors, molecular mass standards for SDS-PAGE, DMSO, TX-100, Histopaque, Sodium dodecyl sulphate (SDS), 5,6,6-tetrachloro-1,1,3,3-tetraethylbenzimidazol-carbocyanine iodide (JC-1), carbonyl cyanide 3-chlorophenylhydrazone (CCCP) and propidium iodide (PI) were obtained from Sigma-Aldrich (Munich, Germany). Tween, Sulphuric acid, acrylamide and dithiotreitol were purchased from Carl Roth GmbH, (Karlsruhe, Germany). Ammonium persulfate and N,N,N,N-tetramethylenediamine were obtained from BioRad (Munich, Germany). 3,3’,5,5’-tetramethylbenzidine was purchased from eBioscience Inc. (San Diego, USA). Following primary antibodies were used: caspase-3, poly (ADP-ribose) polymerase (PARP), claspin, survivin, bcl-2, cytochrome c (Cell Signaling Technology, Danvers, USA); p53 (Santa Cruz biotechnology, Santa Cruz, CA, USA); X-chromosome-linked IAP (XIAP) and Annexin V-APC (BD Bioscience, Heidelberg, Germany); ß-actin-peroxidase antibody (Sigma-Aldrich, Munich, Germany). The Cytotoxicity Detection Kit was purchased from Roche (Grenzach-Wyhlen, Germany).
9
Western Blot Analysis of Apoptosis Markers
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Samples (30 μg) were electrophoresed in sodium dodecyl sulfate (SDS)-polyacrylamide gels, which were transferred to nitrocellulose membranes and probed for the following proteins: caspase 8 (mouse monoclonal antibody (mAb), 1:1,000 dilution, Cell Signaling Technology), caspase 9 (rabbit polyclonal 1:1000 dilution, Cell Signaling Technology), caspase 3 (mouse mAb 1:1,000 dilution, Cell Signaling Technology), poly ADP ribose polymerase (PARP) (rabbit polyclonal 1:1,000 dilution, Cell Signaling Technology), and Tubulin (mouse mAb 1:1,000 dilution, Sigma Aldrich). IRDye 800CW or IRDye 680RD secondary antibodies (1:10,000 dilutions) were used to visualize protein on the Odyssey Imaging system (LI-COR). Protein band pixel density was quantitated (Image Studio version 3.1 software) and the ratio of the cleaved protein band compared to the Tubulin loading control band was used to compare across conditions.
10
Mesothelioma Cell Line Cytotoxicity Assay
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Resveratrol, clofarabine, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrozolum bromide (MTT), phosphate buffered saline (PBS), and antibody to β-actin were obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). Anti-human Mcl-1, Bcl-xL, caspase-3, cleaved caspase-3, and poly (ADP-ribose) polymerase (PARP) antibodies were from Cell Signaling Technologies (Beverly, MA, USA). Trizol reagent, cell culture media and reagents were purchased from Invitrogen (Carlsbad, CA, USA). The human mesothelioma cell line MSTO-211H cells were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum, 1 mM glutamine, 100 units of penicillin/ml and 100 μg of streptomycin/ml. The human normal mesothelial cell line MeT-5A cells were maintained in M199 (Welgene, Daegu, Korea) medium supplemented with 3.3 nM epidermal growth factor, 10% fetal bovine serum, 100 units of penicillin/ml and 100 μg of streptomycin/ml. Cells were grown to 50% confluence in a monolayer culture in this medium for 24 h before treatment.
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