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Cleaved parp

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Cleaved PARP is a lab equipment product that detects the presence of cleaved Poly(ADP-ribose) Polymerase (PARP), a protein involved in cellular processes. This product is used for research purposes and its core function is to provide a tool for the identification and analysis of cleaved PARP.

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Lab products found in correlation

Cleaved caspase 3, by Abcam (23 mentions) Bcl 2, by Abcam (16 mentions) Bcl2 associated x (bax), by Abcam (12 mentions) Gapdh, by Abcam (10 mentions) Cleaved caspase 3, by Cell Signaling Technology (9 mentions) β actin, by Abcam (8 mentions) Caspase 3, by Abcam (8 mentions) β actin, by Santa Cruz Biotechnology (6 mentions) Cleaved caspase 9, by Abcam (5 mentions) P akt, by Cell Signaling Technology (4 mentions) Cyclin d1, by Cell Signaling Technology (4 mentions) Beclin 1, by Abcam (4 mentions) Vimentin, by Abcam (4 mentions) Pvdf membrane, by Merck Group (3 mentions) Caspase 3, by Cell Signaling Technology (3 mentions) Cd133, by Cell Signaling Technology (3 mentions) E cadherin, by Santa Cruz Biotechnology (3 mentions) Bca protein assay kit, by Beyotime (3 mentions) Ripa lysis buffer, by Beyotime (3 mentions) Gapdh, by Cell Signaling Technology (3 mentions)

Spelling variants of this product

Anti-cleaved PARP (19 citations) Anti-Cleaved-parp antibody (4 citations) Cleaved PARP antibody (3 citations) Cleaved poly(ADP‐ribose) polymerase (PARP; (3 citations) Cleaved poly-ADP-ribose polymerase (c-PARP) (2 citations) Cleaved poly-ADP-ribose polymerase (cleaved PARP, (2 citations)

73 protocols using cleaved parp

1

Western Blot Analysis of ROS1 and EMT Markers

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The cells were lysed in RIPA buffer (Beyotime, Haimen, China) containing 1% protease inhibitor PMSF (Beyotime) and centrifuged at 12,000 rpm for 10 min. The supernatant containing total proteins was aspirated and the protein concentration was determined. The total proteins were separated by SDS-PAGE (Beyotime) and then transferred onto PVDF membranes (EMD Millipore, Billerica, MA, USA). After blocking, the membranes were incubated with primary antibodies against ROS1 (1:500, Sangon Biotech, Shanghai, China), cleaved-caspase-3 (1:1000, Abcam, Cambridge Science Park, Cambridge, UK), Bcl-2 (1:400, BOSTER, Wuhan, China), Bax (1:400, BOSTER), cleaved-PARP (1:1000, Abcam), E-cadherin (1:400, BOSTER), Vimentin (1:500, BIOSS, Beijing, China), N-cadherin (1:400, BOSTER), p-PI3K (1:500, BIOSS), PI3K (1:400, BOSTER), p-Akt (1:200, Santa Cruz Biotechnology, Dallas, Texas, USA) and Akt (1:200, Santa Cruz Biotechnology) at 4°C overnight. After washing with TBS-Tween 20 buffer, the membranes were incubated with goat anti-rabbit IgG-HRP (Beyotime) at 37°C for 45 mins. The bands were developed using ECL solution (Beyotime).
Qiao J., Li M., Sun D., Li W, & Xin Y. (2019). Knockdown of ROS proto-oncogene 1 inhibits migration and invasion in gastric cancer cells by targeting the PI3K/Akt signaling pathway. OncoTargets and therapy, 12, 8569-8582.
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2

Immunoblotting and Immunofluorescence Analysis of Autophagy Markers

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Commercial antibodies include: phospho-Histone H3 (Cell Signaling Technology (CST) 9701), cleaved caspase-3 Alexa Fluor 488 (CST 9669S), phospho-p44/42 MAPK (P-ERK) (Invitrogen 44–680G), p44/42 MAPK (ERK1/2) (Invitrogen 13–6200), ATG12 (CST 2010), ATG7 (Santa Cruz Biotechnology sc8668), p62 (Progen Biotechnik GP62C), tubulin (Sigma T6199), p53 (DO1, Calbiochem OP43), cleaved PARP (CST 9451), G6PD (Abcam ab993) LC3 5F10 for IF (Axxora NT0231-00). For immunobloting, we utilized an LC3 antibody which has been described previously and is now commercially available (EMD Millipore ABC232) (35 (link)). Chemicals include: chloroquine (CQ), quinacrine (Q), 6-aminonicotinamide (6-AN), N-acetyl Cysteine (NAC), etoposide (Et) and doxorubicin (DX), all from Sigma-Aldrich, staurosporine (STS, EMD Chemicals), 1-13C glucose (Cambridge Isotopes), 1-14C glucose and 6-14C glucose (Perkin Elmer), and Hoechst (Invitrogen).
Salas E., Roy S., Marsh T., Rubin B, & Debnath J. (2015). Oxidative Pentose Phosphate Pathway Inhibition Is A Key Determinant of Antimalarial Induced Cancer Cell Death. Oncogene, 35(22), 2913-2922.
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3

Antibody Dilution Protocol for Western Blot

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This experiment was carried out as described in our previous study [25 (link)]. The antibodies’ detailed information in this work was listed as below: ALDH1A1 (Cat # 15,910–1-AP, Proteintech; Dilution rate: 1:1000), STARD13 (Cat # AP19692c, Abgent, Wuxi, China; Dilution rate: 1:1000), Oct3/4 (octamer-binding transcription factor 4) (Cat # wl01728, Wanleibio, Shenyang, China; Dilution rate: 1:2000), Ki67 (Cat # ab16667, Abcam; Dilution rate: 1:3500), Cleaved PARP (poly ADP-ribose polymerase) (Cat # ab32064, Abcam; Dilution rate: 1:2000), PARP (Cat # 13,371–1-AP, Proteintech; Dilution rate: 1:1500) and GAPDH (Cat # YFMA0037, YI FEI XUE Biotechnology, Nanjing, China; Dilution rate: 1:5000).
Li X., Jia Q., Zhou Y., Jiang X., Song L., Wu Y., Wang A., Chen W., Wang S, & Lu Y. (2022). Tanshinone IIA attenuates the stemness of breast cancer cells via targeting the miR-125b/STARD13 axis. Experimental Hematology & Oncology, 11, 2.
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4

Antibody and Reagent Characterization Protocol

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The following antibodies were purchased against: xCT (1:1000; cat. no. ab37185; Abcam; Cambridge, UK), cleaved-PARP (1:1000; cat. no. 9541; Cell Signaling Technology; Danvers, MA, USA), GPx4 (1:1000; cat. no. ab125066; Abcam; Cambridge, UK), cystathionine gamma-lyase (CGL; 1:1000; cat. no. 12217-1-AP; Proteintech; San Antonio, TX, USA); GCLC (1:1000; cat. no. GTX16315; Gene Tex, Inc.; San Antonio, TX, USA), β-actin (1:10,000; cat. no. A2228; Sigma-Aldrich; Tokyo, Japan), and cleaved caspase-3 (1:400; cat. no. 9661; Cell Signaling Technology; Danvers, MA, USA).
The following reagents were purchased and dissolved in DMSO to prepare the stock solutions: 100 mM SAS, 10 µM paclitaxel, 10 mM cisplatin, 10 mM 2′,7′-dichlorofluorescin diacetate (DCFH-DA), 1 M propargylglycine (PAG) and 10 mM ferrostatin-1 (all dissolved in DMSO and from Sigma-Aldrich; Tokyo, Japan). A total of 10 mM Z-VAD-FMK (Peptide Institute, Inc.; Osaka, Japan) was dissolved in DMSO. The BODIPY C11 [4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid] was purchased from Thermo Fisher Scientific, Inc.
Idei U., Ohta T., Yamatani H., Seino M, & Nagase S. (2023). Mechanism of Cell Death by Combined Treatment with an xCT Inhibitor and Paclitaxel: An Alternative Therapeutic Strategy for Patients with Ovarian Clear Cell Carcinoma. International Journal of Molecular Sciences, 24(14), 11781.
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5

Western Blot Analysis of Cell Lysates

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Cell pellets were collected and resuspended in 2x Laemmli buffer according to the pellet size. Subsequently probes were sonicated for 15 cycles (30 sec on/ 30 sec off) at high setting using a Diagenode Bioruptor plus. After boiling for 20 min at 95° C, samples were centrifuged for 15 at RT maximum speed and separated on 10% SDS-PAGE for Western Blot analysis. ZRF1 (Homemade), H2B (#12364-Cell Signaling), α-Tubulin (AB15708-Sigma), cleaved PARP (ab32064-Abcam), H2B (#12364-Cell Signaling)AKT (#9272S-Cell Signaling) and P-AKT (#9271T-Cell Signaling) antibodies were used to detect the respective proteins. Relative band intensities were calculated by using the Image Lab program from Bio-Rad.
Kaymak A., Sayols S., Papadopoulou T, & Richly H. (2018). Role for the transcriptional activator ZRF1 in early metastatic events in breast cancer progression and endocrine resistance. Oncotarget, 9(47), 28666-28690.
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6

Antibody-Based Protein Analysis of Cellular Pathways

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The following antibodies were purchased against: xCT (1:1,000; cat. no. ab37185; Abcam), cleaved-PARP (1:1,000; cat. no. 9541; Cell Signaling Technology, Inc.), Akt (1:1,000; cat. no. 9272; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (1:1,000; cat. no. 4058; Cell Signaling Technology, Inc.), p44/42 MAPK (Erk1/2; 1:1,000; cat. no. 4695; Cell Signaling Technology, Inc.), p-p44/42 MAPK (1:1,000; cat. no. 9106; Cell Signaling Technology, Inc.), p-SAPK/JNK (1:1,000; cat. no. 9251; Cell Signaling Technology, Inc.), JNK2 (1:200; cat. no. sc7345; Santa Cruz Biotechnology, Inc.) and β-actin (1:1,000; cat. no. A2228; Sigma-Aldrich; Merck KGaA).
The following reagents were purchased and dissolved in DMSO or distilled water to prepare the stock solutions: 100 mM SAS, 10 mM 2′,7′-dichlorofluorescin diacetate (DCFH-DA) and 10 mM ferrostatin-1 (all dissolved in DMSO and from Sigma-Aldrich; Merck KGaA). A total of 10 mM deferoxamine mesylate (DFO) (cat. no. ab120727; Abcam) was dissolved in water and 10 mM Z-VAD-FMK (Peptide Institute, Inc.) in DMSO.
Sugiyama A., Ohta T., Obata M., Takahashi K., Seino M, & Nagase S. (2020). xCT inhibitor sulfasalazine depletes paclitaxel-resistant tumor cells through ferroptosis in uterine serous carcinoma. Oncology Letters, 20(3), 2689-2700.
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7

Histological Analysis of U87 MG Tumor

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For histological analysis, the mice were sacrificed after 44 days of the U87 MG efflux cell injection. Cardiac perfusion was performed and frozen blocks were prepared for sectioning as previously reported [54 (link)]. The tissues were stained with hematoxylin and eosin (H&E) to mark the tumor volume within the sections. Immunofluorescence was performed using primary antibodies as follows: p-p38 (1:400, Cell Signaling), cleaved caspase-3 (1:100, Abcam), survivin (1:250, Abcam), and cleaved PARP (1:400, Abcam). The secondary antibody used was Alexa Fluor 594- or 488-conjugated with anti-IgG or 594-conjugated antigoat IgG (1:500; Invitrogen, Carlsbad, CA, USA), and the sections were mounted with anti-fading solution containing 4′-6-diamidino-2-phenylindole (DAPI, Vector Laboratories, Burlingame, CA, USA). Fluorescent images were obtained using a fluorescence microscope (Leica, DMi8, Wetzlar, Germany). Positively stained slides were quantified from a minimum of three randomly stained slides.
Akter M., Jangra A., Choi S.A., Choi E.H, & Han I. (2020). Non-Thermal Atmospheric Pressure Bio-Compatible Plasma Stimulates Apoptosis via p38/MAPK Mechanism in U87 Malignant Glioblastoma. Cancers, 12(1), 245.
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8

Western Blot Analysis of Cellular Proteins

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Protein lysates (30 µg/lane) were separated on 10% SDS-PAGE gels and transferred to PVDF membrane. Following blocking, target proteins were detected by probing overnight at 4°C with antibodies against: NFIB, Bin1 and p19ARF (Abcam); PARP, cleaved PARP, cleaved caspase-3, caspase-3, cyclin D1, CDK4, CDK6, p-Akt-ser473, β-catenin, and CD133 (Cell Signaling Technologies); and total Akt, p-Erk and ERK (Santa Cruz Biotechnology). Then, membranes were washed and incubated with anti-mouse/rabbit IgG secondary antibody (Invitrogen) conjugated with horseradish peroxidase (HRP) at room temperature. Specific proteins were visualized using an enhanced chemiluminescence detection reagent (Pierce; ThermoFisher Scientific, Inc.).
Perumal N., Kanchan R.K., Doss D., Bastola N., Atri P., Chirravuri-Venkata R., Thapa I., Vengoji R., Maurya S.K., Klinkebiel D., Talmon G.A., Nasser M.W., Batra S.K, & Mahapatra S. (2021). MiR-212-3p functions as a tumor suppressor gene in group 3 medulloblastoma via targeting nuclear factor I/B (NFIB). Acta Neuropathologica Communications, 9, 195.
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9

Autophagy and EMT Regulation Pathways

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Chloroquine (CQ), 3-methyladenine (3-MA), rapamycin and TGF- β2 were purchased from Sigma-Aldrich (St Louis, MO, USA). LY2109761 was from Selleck Chemicals (Houston, TX, USA). Antibodies against microtubule-associated protein 1 light chain 3-I (LC3-I), LC3-II, p62, Beclin 1, ATG7, poly (ADP-ribose) polymerase (PARP), cleaved-PARP, caspase 9, cleaved-caspase 9, Bax, and Bcl-x1 were from Abcam (Cambridge, MA, USA). Antibodies against TGF-β2, Smad2, p-Smad2, E-cadherin, α-catenin, β-catenin, N-cadherin, fibronectin, ZO-1, Vimentin, MMP-9, Snail, Slug and β-actin were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Primary antibodies of LC3-II and E-cadherin and secondary antibodies Alexa Fluor 568 anti-mouse IgG and Alexa Flour 568 anti-rabbit antibodies were from Jackson Immuno Research (Lancaster, PA, USA).
Li Z., Lu C., Wang F., Guo H., Wang Z., Yin H, & Li J. (2023). Heat treatment-induced autophagy promotes breast cancer cell invasion and metastasis via TGF-β2-mediated epithelial-mesenchymal transitions. PeerJ, 11, e14640.
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10

Western Blotting of Tumor Cell Lysates

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Western blotting was performed as previously described4 (link) with slight modifications: unless otherwise noted, protein lysates from tumor cells were extracted using 1X Cell Lysis Buffer (CST 9803) supplemented with PhosSTOP phosphatase inhibitors (Sigma) and cOmplete™, Mini EDTA-free Protease Inhibitor Cocktail (Sigma), and samples were normalized by total protein concentration measured by the Bio-Rad Protein Assay Dye Reagent (Bio-Rad 5000006) or equal numbers of nuclei, as indicated.
Primary antibodies used include cleaved-PARP (CST 9541), Bcl-xL (Abcam ab32370), RB (CST 9309), Lamin A/C (CST 4777), JunB (CST 3753), c-Jun (CST 9165), Fra-2 (CST 19967) and vinculin (Sigma V9131). Primary antibodies were used at 1:1,000, except vinculin, which was used at 1:5,000. Secondary antibodies used include Quick Western Kit IRDye 680RD (Licor 926–68100) used at 1:1000, Mouse IgG (H&L) Antibody DyLight™ 800 Conjugated (Rockland 610–145-002) used at 1:5,000, and Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 680 (Invitrogen A21109) used at 1:3,000. Secondary antibodies with different lot numbers were used and consistent results were observed. Western blot images were acquired on the Odyssey CLx Imaging System (LI-COR Biosciences) and processed using ImageStudio Lite v5.2.5 software.
Watt A.C., Cejas P., DeCristo M.J., Metzger-Filho O., Lam E.Y., Qiu X., BrinJones H., Kesten N., Coulson R., Font-Tello A., Lim K., Vadhi R., Daniels V.W., Montero J., Taing L., Meyer C.A., Gilan O., Bell C.C., Korthauer K.D., Giambartolomei C., Pasaniuc B., Seo J.H., Freedman M.L., Ma C., Ellis M.J., Krop I., Winer E., Letai A., Brown M., Dawson M.A., Long H.W., Zhao J.J, & Goel S. (2020). CDK4/6 inhibition reprograms the breast cancer enhancer landscape by stimulating AP-1 transcriptional activity. Nature cancer, 2(1), 34-48.
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