Cleaved parp

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Cleaved PARP is a lab equipment product that detects the presence of cleaved Poly(ADP-Ribose) Polymerase (PARP), a protein that plays a role in DNA repair and cell death processes. This product provides a reliable method for measuring PARP cleavage, which is a widely used indicator of apoptosis or programmed cell death.

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Lab products found in correlation

Close spelling variants of this product

Anti-cleaved PARP (17 citations) Cleaved PARP-1 (12 citations) Anti-cleaved PARP-1 (6 citations)

42 protocols using cleaved parp

Cinobufagin-Induced Apoptosis and Autophagy

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Cinobufagin was obtained from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China) and dissolved in Dulbecco's modified Eagle's medium (DMEM) (Gibco, Grand Island, NY, USA) to each working dose. Fetal bovine serum (FBS) was purchased from Hangzhou Sijiqing Biological Engineering Material Co., Ltd. (Hangzhou, Zhejiang, China). Chloroquine (CQ), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), N-acetyl cysteine (NAC) and 3-methyladenine (3-MA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The Annexin V-FITC Apoptosis Detection Kit I was obtained from BD Biosciences (San Diego, CA, USA). Hoechst 33258 was purchased from Promega Corp. (Madison, Wi, USA). Lipofectamine 2000 transfection reagent was purchased from Invitrogen Life Technologies (Carlsbad, CA, USA). Antibodies against caspase-3, caspase-8, caspase-9, cleaved PARP, phospho-JNK, total JNK, phospho-p38, total p38, LC3B, p62, GAPDH, enhanced chemiluminescence (ECL) and western blotting kits were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

MA K., ZHANG C., HUANG M.Y., LI W.Y, & HU G.Q. (2016). Cinobufagin induces autophagy-mediated cell death in human osteosarcoma U2OS cells through the ROS/JNK/p38 signaling pathway. Oncology Reports, 36(1), 90-98.

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Lung Cancer Cell Line Characterization

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NSCLC cell lines A549, HCC4006 and Calu-1 (three human lung adenocarcinoma cell lines), and SK-MES-1 and KLN205 (two human lung squamous carcinoma cell lines) were obtained from the Shanghai Institute for Biological Science (China). All cell lines were purchased between 2012 and 2015 and authenticated based on growth rate, morphology, and viability and were frequently confirmed to be mycoplasma free. The cells were cultured in RPMT 1640 media supplemented with 10% heat-inactivated fetal calf serum (FBS), L-glutamine and 100 U/ml penicillin, and 100 μg/ml streptomycin sulfate. The cells were maintained in a humidified incubator in 5% CO₂ at 37 °C.
Antibodies to various antigens were as follows: CHK1, PCHK1 (S296), PCHK1 (S317), PCHK1 (S345), CHK2, PCHK2 (T68), PCHK2 (S516), Cdc25A, PCdc25C (S216), H2AX, γH2AX (S139), PH3 (S10histone), ATM, PATM (S1981), DNA-PKCs, PDNA-PKCs (S2056), and GAPDH were from Cell Signaling. FANCL, FANCD2, BRCA2, PARP1, RAD51, caspase-3, cleaved caspanse-3, PARP and cleaved PARP from Santa Cruz. Gemcitabine was from Hanson Pharmaceutic (China), Cisplatin from Yangtze River Pharmaceutic (China), MK-8776 from Selleck Chemicals, CHK2 inhibitor II from Abcam. Gemcitabine was dissolved in PBS. Cisplatin, MK-8776 and CHK2 inhibitor II were dissolved in DMSO and used at the specified concentrations.

Dai C.H., Wang Y., Chen P., Jiang Q., Lan T., Li M.Y., Su J.Y., Wu Y, & Li J. (2017). Suppression of the FA pathway combined with CHK1 inhibitor hypersensitize lung cancer cells to gemcitabine. Scientific Reports, 7, 15031.

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Apoptosis Regulation by CTD Compound

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CTD (purity ≥98%, product number: MB6525) was purchased from Dalian Meilun Biotechnology Co., Ltd. and added with an appropriate amount of DMSO solution to dissolve into a 32 mM stock solution, stored at −20°C for further use. The antibodies against caspase 3, cleaved caspase 3, cleaved caspase 8, Cyclin E, Cyclin B1, CDK2, p21, p27, and p53 were obtained from Wanlei Biotechnology (Shenyang, China). The antibodies against Nur77, PARP, cleaved PARP, c-Jun, Jun-B, Bcl-2, and Bax were purchased from Santa Cruz Biotechnology (CA, USA). The antibodies against β-actin, mouse, or rabbit IgG were obtained from Sigma-Aldrich (St. Louis, MO, USA). Hoechst 33342 was obtained from Wanlei Biotechnology (Shenyang, China). Other chemicals were obtained from Sigma-Aldrich, unless indicated otherwise.

Yu Z., Li L., Wang C., He H., Liu G., Ma H., Pang L., Jiang M., Lu Q., Li P, & Qi H. (2020). Cantharidin Induces Apoptosis and Promotes Differentiation of AML Cells Through Nuclear Receptor Nur77-Mediated Signaling Pathway. Frontiers in Pharmacology, 11, 1321.

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Western Blot Analysis of NF-κB Pathway

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PANC-1 and BxPC-3 cells (treated with the test compound after 24 h starvation, and then stimulated with 1 ng/mL TNF-α for 30 min) were lysed in a lysis buffer for 15 min on ice. The cell lysates were centrifuged at 4°C to remove insoluble components. Protein (80 μg) from each sample was subjected to SDS-PAGE and transferred onto PVDF membranes (Bio-Rad Laboratories). After incubation with 5% skim milk in TBST for 1.5 h, the membranes were incubated with primary antibodies (pIKKβ, IκB, cleaved PARP, bcl-2, and GAPDH; Santa Cruz) at 4°C overnight and then with the horseradish peroxidase-labeled secondary antibody at room temperature for 1 h. The results were visualized using enhanced chemiluminescence reagents (Bio-Rad Laboratories), and the band densities were quantified using ImageJ analysis software (National Institutes of Health, Bethesda, MD, USA).

Xie X., Tu J., You H, & Hu B. (2017). Design, synthesis, and biological evaluation of novel EF24 and EF31 analogs as potential IκB kinase β inhibitors for the treatment of pancreatic cancer. Drug Design, Development and Therapy, 11, 1439-1451.

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Protein Expression Analysis by Western Blot

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Cells were harvested and homogenized in RIPA lysis buffer and centrifuged at 12,000 rpm for 20 min at 4°C. Protein concentrations of the supernatants were determined using BCA Protein Quantification Kit (Vazyme biotech co., Itd.). Western blot analysis was performed as previously described [38 (link)], using rabbit polyclonal antibodies to human survivin and c-IAP1 (R&D systems, MN), XIAP and Cyclin B1 (Cell signaling, MA), phosphor-Cdc25C, Cdc2, phospho-Cdc2, cleaved PARP and cleaved caspase-3 (Santa Cruz, CA).

Qin Q., Cheng H., Lu J., Zhan L., Zheng J., Cai J., Yang X., Xu L., Zhu H., Zhang C., Liu J., Ma J., Zhang X., Dai S, & Sun X. (2014). Small-molecule survivin inhibitor YM155 enhances radiosensitization in esophageal squamous cell carcinoma by the abrogation of G2 checkpoint and suppression of homologous recombination repair. Journal of Hematology & Oncology, 7, 62.

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Mitochondrial Dysfunction and Muscle Atrophy

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t-BHP, FCCP, antimycin A, oligomycin and ATP assay kit were from Sigma (St. Louis, MO, USA). Fetal bovine serum (FBS) was from PAA Laboratories GmbH (Linz, Austria). High glucose Dulbecco's modified Eagle's medium (DMEM), horse serum (HS), JC-1 (5,5,6,6-tetrachloro-1,1,3,3-tetraethylbenzimidazolylcarbocyanine iodide), DAPI and antibodies against complex I, II, III, IV and V were from Invitrogen (Carlsbad, CA, USA). HRP-conjugated anti-mouse/rabbit IgG antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). The antibodies against MyHC, AFG312, PARP and cleaved PARP were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The antibodies against β-actin, cleaved caspase 3 and cleaved caspase 9 were from Cell Signaling Technology (Danvers, MA, USA). The antibody against OPA1 was from BD (Franklin Lakes, NJ, USA). OPA1 siRNA oligo and PCR primers for Afg3l2, MyHC I, MyHC IIb, MyHC IIx, Murf-1, Atrogin-1 and β-actin were synthesized by Genepharma (Shanghai, China).

Wang X., Li H., Zheng A., Yang L., Liu J., Chen C., Tang Y., Zou X., Li Y., Long J., Liu J., Zhang Y, & Feng Z. (2014). Mitochondrial dysfunction-associated OPA1 cleavage contributes to muscle degeneration: preventative effect of hydroxytyrosol acetate. Cell Death & Disease, 5(11), e1521-.

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Molecular Mechanisms of MGCD-Induced Apoptosis

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MGCD was kindly provided by ROCHE R & D CENTER (CHINA) LTD. Nutlin-3 was purchased from Sigma. For western blot detection, we used Ac-Histone H3 (Upstate), Histone H3, Cleaved caspase-3, Bcl-2, Cyclin B1, Bax (Epitomics), p-cdc2 (Tyr15), cdc2, p53, p21 (Cell Signaling Technology), cleaved PARP (Santa Cruz), and GAPDH, tubulin (Proteintech Group) antibodies.

Yan M., Qian Y.M., Yue C.F., Wang Z.F., Wang B.C., Zhang W., Zheng F.M, & Liu Q. (2016). Inhibition of histone deacetylases induces formation of multipolar spindles and subsequent p53-dependent apoptosis in nasopharyngeal carcinoma cells. Oncotarget, 7(28), 44171-44184.

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Evaluating Docetaxel and Bicalutamide Sensitivity

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Docetaxel (CAS#114977-28-5) was purchased from TSZ CHEM (Framingham, MA). Bicalutamide (Cat: #B3209) were purchased from LKT Laboratories, Int. Elacridar (Cat: #143664-11-3) and Rhodamine 123 (Cat: #62667-70-9) were purchased from Sigma-Aldrich. Anti-ser15 phosphorylated p53 antibody was obtained from Cell Signaling Technology, Int. Cleaved PARP, wt-p53 and tubulin antibodies were obtained from Santa Cruz Biotechnologies (Santa Cruz, CA).

Zhu Y., Liu C., Armstrong C., Lou W., Sandher A, & Gao A.C. (2015). Anti-androgens inhibit ABCB1 efflux and ATPase activity and reverse docetaxel resistance in advanced prostate cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 21(18), 4133-4142.

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Western Blot Analysis of Cellular Markers

Cited 1 time

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Western blotting was conducted as previously described 34 (link). The following primary antibodies were used in this study: GRP78 (# 3177; Cell Signaling Technology), GRP94 (#3196-1; Epitomics), pERK1/2 (#sc-377400, Santa Cruz Biotechnology), p65 (#sc-8008; Santa Cruz Biotechnology), CHOP(#2895; Cell Signaling Technology), cleaved PARP (#sc-56196, Santa Cruz Biotechnology), NLRP3 (#sc-134306; Santa Cruz Biotechnology), HXK2 (#sc-374091, Santa Cruz Biotechnology), IL-1β (#sc-12742; Santa Cruz Biotechnology), IL-18 (#sc133127; Santa Cruz Biotechnology), p-eIF2a (#33981; Cell Signaling Technology), Casp1 (#YH050707C; Epitonics), ZDHHC1 (#AP10315a; Abgent), E-cadherin (#sc8426; Santa Cruz Biotechnology), Occludin (#TA306787; Origene), N-cadherin (#610920; BD transduction laboratories), Vimentin (#5741s; Cell Signaling Technology), G6PD (#12263s; Cell Signaling Technology), Casp3 (#9664; Cell Signaling Technology), Casp7 (#8438; Cell Signaling Technology), GLUT1(#sc-377228; Santa Cruz Biotechnology), beta-actin (#sc8432; Santa Cruz Biotechnology), GAPDH (#sc-47727; Santa Cruz Biotechnology).

Le X., Mu J., Peng W., Tang J., Xiang Q., Tian S., Feng Y., He S., Qiu Z., Ren G., Huang A., Lin Y., Tao Q, & Xiang T. (2020). DNA methylation downregulated ZDHHC1 suppresses tumor growth by altering cellular metabolism and inducing oxidative/ER stress-mediated apoptosis and pyroptosis. Theranostics, 10(21), 9495-9511.

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Anticancer Activity of Compound Cocktail

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Methyl alcohol (product no. 62), n-hexane (product no. 4794), ethyl acetate (product no. 2936), and chloroform (product no. 1268) were purchased from Duksan chemicals, South Korea. 1-butanol (cat# 33065) was purchased from Honeywell research chemicals. 5-Fluorouracil (cat# F6627), propidium iodide (PI), and 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Dihydrorhodamine 123 (DHR 123: cat# D23806) was purchased from ThermoFisher scientific. Antibodies [cleaved caspase-3 (cat# sc-22171), cleaved PARP (cat# sc-56196), and β-actin (cat# sc-47778)] were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). 4′,6-diamidino-2-phenylindole (DAPI) mounting solution (Cat# H-1200) was purchased from Vector laboratories, USA. All the chemicals and reagents were used as directed by the manufacturers.

Islam S.U., Ahmed M.B., Shehzad A, & Lee Y.S. (2019). Methanolic Extract of Artemia salina Eggs and Various Fractions in Different Solvents Contain Potent Compounds That Decrease Cell Viability of Colon and Skin Cancer Cell Lines and Show Antibacterial Activity against Pseudomonas aeruginosa. Evidence-based Complementary and Alternative Medicine : eCAM, 2019, 9528256.

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