Gefitinib (cat. no. S1025) was purchased from Selleck Chemicals (Houston, TX, USA) and CQ (cat. no. A506569) was purchased from Sangon Biotech Co., Ltd. (Shanghai, China). Gefitinib and CQ were dissolved in DMSO and DMEM, respectively, and subsequently stored at a stock concentration of 100 mM at −20°C. The following primary antibodies and dilutions were used in the present study: Microtubule associated protein 1 light chain 3-B (LC3B; cat. no. 3868S; Cell Signalling Technology, Inc., Danvers, MA, USA; 1:1,000), caspase-3 (cat. no. sc-7272; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; 1:1,000), poly-(ADP-ribose) polymerase (PARP; cat. no. 9532S, Cell Signalling Technology, Inc.; 1:5,000), β-actin (cat. no. sc-47778; Santa Cruz Biotechnology, Inc., Inc.; 1:1,000) and α-tubulin (cat. no. sc-5286; Santa Cruz Biotechnology; Inc.; 1:1,000). Secondary antibodies used in the present study were horseradish peroxidase (HRP)-tagged anti-mouse IgG (cat. no. 31430; Invitrogen; Thermo Fisher Scientific, Inc.; 1:5,000) and HRP-tagged anti-rabbit IgG (cat. no. 31460; Invitrogen; Thermo Fisher Scientific, Inc.; 1:5,000).
Poly adp ribose polymerase parp
Manufactured by Santa Cruz Biotechnology
Sourced in United States, Canada
Poly (ADP-ribose) polymerase (PARP) is an enzyme that catalyzes the addition of poly(ADP-ribose) chains to target proteins. It plays a role in various cellular processes, including DNA repair, gene regulation, and cell death.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by Santa Cruz Biotechnology (7 mentions)
Caspase 3, by Santa Cruz Biotechnology (7 mentions)
Propidium iodide, by Merck Group (6 mentions)
Bcl 2, by Santa Cruz Biotechnology (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (4 mentions)
Bcl xl, by Santa Cruz Biotechnology (4 mentions)
α tubulin, by Santa Cruz Biotechnology (3 mentions)
Caspase 3, by Cell Signaling Technology (3 mentions)
Anti pan cadherin, by Thermo Fisher Scientific (3 mentions)
Anti pten, by Thermo Fisher Scientific (3 mentions)
Anti p pten ser 380 thr 382 ser 385, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions)
β actin, by Cell Signaling Technology (2 mentions)
Caspase 9, by Santa Cruz Biotechnology (2 mentions)
P perk, by Santa Cruz Biotechnology (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Simvastatin, by Merck Group (2 mentions)
35 protocols using poly adp ribose polymerase parp
1
Gefitinib and Chloroquine Autophagy Inhibition
2
Western Blot Analysis of Protein Expression
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HeLa or PK-15 cells were lysed in the precooled lysis buffer added with phenylmethylsulfonyl fluoride (PMSF; Beyotime, China). The samples were resolved in a 12% sodium dodecyl sulfate-polyacrylamide gel. Separated proteins were then transferred onto a nitrocellulose membrane and incubated with antibody against STAT1 (Cell Signaling, Danvers, MA, USA), phospho-STAT1 (Tyr701, herein named STAT1-Y701, Cell Signaling), FLAG (Sigma-Aldrich, St. Louis, MO, USA), HA (Sigma-Aldrich), heat shock protein 90 (HSP90; Santa Cruz Biotechnology, Santa Cruz, CA, USA), poly (ADP-ribose) polymerase (PARP; Santa Cruz Biotechnology), rabbit anti-3C serum (kept in our laboratory) or β-actin antibody (Santa Cruz Biotechnology), respectively. The membranes were then incubated with HRP-conjugated affinipure goat anti-rabbit IgG or goat anti-mouse IgG (Boster, Wuhan, China), respectively. The membranes were then developed using WesternBright™ Sirius detection kit on the basis of the manufacturer's instructions (Advansta, Menlo Park, CA, USA). Digital signal was acquired and analyzed by the Quantity One program, version 4.6 (Bio-Rad).
3
Apoptosis Pathway Evaluation Protocol
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RPMI-1640, streptomycin, penicillin G, 3-(4, 5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), TA, Rhodamine-123 and 2,7-Dichlorodihydrofluorescein diacetate (DCFH-DA) were obtained from Sigma-Aldrich, Merck KGaA (Darmstadt, Germany). Foetal bovine serum (FBS) was obtained from Gibco (Thermo Fisher Scientific, Inc.,Waltham, MA, USA). Pro caspase 3, caspase 9, poly (ADP-ribose) polymerase (PARP), β actin and Annexin V/propidium iodide (PI) was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).
4
Evaluating Cellular Stress Responses
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Propidium iodide (PI), N-acetyl-L-cysteine (NAC), DCFH-DA (6-carboxy-2,7-dichlorodihydrofluorescein diacetate), and PEITC were purchased from Sigma Aldrich (St. Louis, MO, USA) and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) from Amresco (Solon, OH, USA). Fluorescein isothiocyanate (FITC)-labeled annexin V (Annexin V-FITC) kit was obtained from BD Biosciences Pharmingen (San Diego, CA, USA). Antibodies against ATF-6 and IRE1α were purchased from Cell Signaling Technology (CA), P-PERK, CHOP/GADD153, and poly-ADP ribose polymerase (PARP) from Santa Cruz Biotechnology (CA), BiP/GRP78 from BD Transduction LaboratoriesTM, and GAPDH from AbFrontier (Seoul, South Korea).
5
Antibody Characterization for Cell Signaling
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Antibodies against caspase-3, caspase-9, and caspase-12 were obtained from Calbiochem (San Diego, CA, USA). Anti- p85α -Akt, -phospho (p)-Akt (Ser 473), BAX, BAK, BCL-2, BCL-XL, and cytochrome c (Cyt c) antibodies were purchased from BD PharMingen. Antibodies directed against acid sphingomyelinase (ASM), BAD, p-BAD (Ser 128), p-BCL-2 (Ser 87), Bid, calnexin, p-CDK1 (Thr 161), Cyt c oxidase subunit II (Cox-2), and MCL-1 were obtained from Abcam (Cambridge, MA, USA). Antibodies against CD55, CD71, nucleolin, p21, CDK1, CDK2, cyclin A, cyclin B1, cyclin D, cyclin E, p110α, poly (ADP-ribose) polymerase (PARP), and Rac1 were purchased from Santa Cruz Biotechnology. Anti-pan-cadherin, anti-PTEN, and anti-p-PTEN (Ser 380/Thr 382/Ser 385) antibodies were obtained from Thermo Fisher Scientific (New York, NY, USA). Antibodies against β-actin, p-BCL-2 (Thr 69), p-BCL-XL (Ser 62), FLAG-epitope tag, and α-tubulin were obtained from Sigma-Aldrich. Horseradish peroxidase (HRP)-conjugated streptavidin was provided from Thermo Fisher Scientific (New York, NY, USA). HRP-conjugated anti-mouse, -goat, and -rabbit IgG secondary antibodies were purchased from Jackson ImmunoResearch Laboratory (West Grove, PA, USA).
6
Glabridin Induces Apoptosis via MAPK Pathways
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Glabridin, ≥98% (HPLC), powder was purchased from Sigma Chemical Co. (St. Louis, MO, USA). Stock solution of Glabridin was made at 10, 20 and 40 µM concentration in DMSO and stored at −20°C. The final concentration of DMSO for all treatments was consistently less than 0.1%. 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) was obtained from Sigma Chemical Co. (St. Louis, MO, USA). Specific inhibitors for ERK1/2 (U0126), JNK1/2 (SP600125) or p38 (SB202190) were purchased from Calbiochem (San Diego, CA). Antibodies, specifically of Bax, Bad, Bid, Bcl-2, cleaved caspase-3, caspase-8, caspase-9, poly (ADP-ribose) polymerase (PARP), p-extracellularly regulated kinase (ERK)1/2, p-p38, p-c-Jun N-terminal kinase (JNK), ERK1/2, p38, JNK1/2, α-tubulin and β-actin (for the Western blot analysis), were purchased from Santa Cruz Biotechnology.
7
Quantification of STIM1 and PARP Protein Expression
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Total protein was extracted using 2X SDS Lysis Buffer (100 mM Tris, 4% SDS, 10% glycerol, 200 mM NaCl, 2 mM EDTA, pH=6.8) from SW1116 and HCT116 cells, and then 30 g protein was separated by SDS-PAGE with 10% acrylamide and transferred onto polyvinylidene fluoride membranes. The membranes were probed with specific antibodies against STIM1 (cat. no. sc-166840; 1:1,000 for HCT116 cells and 1:500 for SW1116 cells; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), poly(ADP-ribose) polymerase (PARP; cat. no. 9542; 1:1,000; Cell Signaling Technology Europe, B.V., Leiden, The Netherlands) or GAPDH (cat. no. 10494-1-AP; 1:50,000 for HCT116 cells and 1:10,000 for SW1116 cells; Proteintech Group Inc., Chicago, IL, USA) at 4°C overnight. Membranes were subsequently incubated with horseradish peroxidase-conjugated goat anti-mouse (cat. no. sc-2005; 1:5,000; Santa Cruz Biotechnology, Inc.) or goat anti-rabbit (cat. no. sc-2054, 1:5,000; Santa Cruz Biotechnology, Inc.) immunoglobulin G antibody and visualized using the Pierce enhanced chemiluminescence kit (Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol.
8
Apoptosis Pathway Analysis Protocol
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RPMI 1640, fetal bovine serum (FBS), and antibiotics were purchased from Gibco-BRL (Grand Island, NY). Sulindac, simvastatin, propidium iodide (PI), 3-(4,5-dimethyl-2- thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), bicinchoninic acid, dimethyl sulfoxide (DMSO), N-acetylcysteine (NAC), and LY294002 were purchased from Sigma-Aldrich (St. Louis, MO). The following primary antibodies were used caspase-3, -8, -9, poly(ADP-ribose) polymerase (PARP; Santa Cruz Biotechnology, Santa Cruz, CA), serine/threonine protein kinase (AKT), phospho-AKT, survivin, X-linked inhibitor of apoptosis protein (XIAP), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Cell Signaling Technology, Beverly, MA). Anti-rabbit IgG-conjugated horseradish peroxidase (HRP) antibodies and an enhanced chemiluminescent (ECL) kit were purchased from Amersham Pharmacia Biotech (Buckinghamshire, UK).
9
Western Blot Analysis of Apoptosis Regulators
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Antibodies against cytochrome c, poly (ADP-ribose) polymerase (PARP), and β-actin were purchased from Santa Cruz Biotechnology. Antibodies against caspase-8, caspase-9, Fas, TNFR1, TNF-α, DR4, DR5, and p53 were purchased from Abcam (Hong Kong). Horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Pierce Biotechnology. Equal amounts of cell lysates were separated by SDS-PAGE and electrotransferred onto immobilon transfer membranes (Millipore, Bedford, MA). The membranes were blocked with 5% skim milk and probed with the indicated antibodies. The blots were washed and incubated with an HRP-coupled anti-mouse or anti-rabbit IgG antibody, followed by detection with ECL-enhanced chemiluminescence detection reagents (Amersham Biosciences, Piscataway, NJ). β-actin, α-tubulin or COXII were used as loading controls.
10
Evaluating Cell Signaling and Proliferation
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Dulbecco’s Modified Eagle Medium (DMEM), trypsin and penicillin-streptomycin were supplied from Gibco (Grand Island, NY, USA). Fetal bovine serum (FBS), RIPA buffer, protease inhibitors and Coomassie Plus™ Protein Assay Reagent were obtained from Thermo Scientific Company (Waltham, MA, USA). Guava Cell Nexin Reagent was purchased from Guava Technologies (Darmstadt, Germany). Nitrocellulose membrane and ECL reagent were supplied from GE Healthcare (Little Chalfont, UK). Gelatin, propidium iodide (PI) and 3-Methyladenine (3-MA) were obtained from Sigma (St. Louis, MO, USA). Antibodies specific to COX-2, and cyclin -D1 were purchased from Millipore (Darmstadt, Germany). Antibodies specific to β-actin, uPA, urokinase-type plasminogen activator receptor (uPAR), poly (ADP-ribose) polymerase (PARP), and p65 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies for the detection of ERK1/2, p38, JNK, c-Jun, and p65 were purchased from Cell Signaling Technology (Danvers, MA, USA). Matrigel was purchased from Becton Dickinson (Bedford, MA, USA).
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