Protein lysates from 48 h co-cultures of GFP-H1299 and ACE2/TMPRSS2+, and Spike-H1299 and ACE2/TMPRSS2+ pretreated with DMSO or 10 μM AI for 1 h were prepared by dissolving the co-cultures directly in 2X SDS sample buffer, separated by SDS-PAGE, and transferred onto nitrocellulose membranes. The membranes were then blocked with 5% skim milk in TBST (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 0.05% Tween 20) for 1 h at RT, followed by incubation with primary antibodies in 1% bovine serum albumin (BSA) in TBST overnight at 4 °C. The primary antibodies used were as follows; rabbit anti-Cleaved Caspase-9 (Cell Signaling, #9505), rabbit anti-Cleaved PARP (Cell Signaling, #9541), and mouse anti-GAPDH (Abcam, ab8245). The membranes were incubated with the corresponding horseradish peroxidase-conjugated secondary antibodies (KPL, USA) for 1 h at RT. Immuno-reactive protein bands were detected using ECL Western Blotting Substrate (Thermo Fisher Scientific, USA).
Rabbit anti cleaved parp
Manufactured by Cell Signaling Technology
Sourced in United States
Rabbit anti-cleaved PARP is a primary antibody that specifically recognizes the cleaved form of Poly(ADP-ribose) Polymerase (PARP) protein. PARP is a key enzyme involved in the DNA damage response, and its cleavage is a hallmark of apoptosis. This antibody can be used to detect and study the activation of programmed cell death pathways.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti cleaved caspase 3, by Cell Signaling Technology (7 mentions)
Mouse anti β actin, by Merck Group (7 mentions)
Rabbit anti cleaved caspase 9, by Cell Signaling Technology (4 mentions)
Rabbit anti parp, by Cell Signaling Technology (3 mentions)
Rabbit anti akt, by Cell Signaling Technology (3 mentions)
Mouse anti β actin, by Santa Cruz Biotechnology (3 mentions)
Mouse anti gapdh, by Santa Cruz Biotechnology (3 mentions)
Rabbit anti cleaved caspase 7, by Cell Signaling Technology (3 mentions)
Mouse anti gapdh, by Abcam (2 mentions)
Rabbit anti gapdh, by Cell Signaling Technology (2 mentions)
Mouse anti α tubulin, by Merck Group (2 mentions)
Rabbit anti bax, by Cell Signaling Technology (2 mentions)
Rabbit anti bcl 2, by Cell Signaling Technology (2 mentions)
Rabbit anti bcl xl, by Cell Signaling Technology (2 mentions)
Rabbit anti p akt, by Cell Signaling Technology (2 mentions)
Rabbit anti mtor, by Cell Signaling Technology (2 mentions)
Bioimager 600 system, by Analytik Jena (2 mentions)
Mouse anti ago2, by Abnova (2 mentions)
Rabbit anti dicer1, by Fortis Life Sciences (2 mentions)
Rabbit anti trbp2, by Cell Signaling Technology (2 mentions)
31 protocols using rabbit anti cleaved parp
1
Quantifying Apoptosis Signaling in SARS-CoV-2 Co-Cultures
2
Tracking Inducible Apoptosis Markers
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To measure changes in the fraction of shRNA-containing mCherry-positive cells, cells were re-plated every 3 or 4 days. At those time points, approximately 25% of the culture was used to measure the percentage of mCherry-positive cells by flow cytometry, whereas the remaining cells were re-plated for further time points. If indicated, cells were treated with doxycycline (1 μg per mL) or ethanol as a solvent control. At least 10,000 (BT-549) or 30,000 (KBM-7) events were analyzed per sample on an LSR-II (Becton Dickinson). Cells, pre-treated with doxycycline (1 μg per mL) or HU, were harvested at different time points, washed and fixed in ice-cold 70% ethanol. Cells were permeabilized and blocked with PBS–1% BSA–0.05% Tween-20 or with PBS–2% BSA–0.1% Triton for 1 h and stained with rabbit anti-cleaved PARP (1:100, Cell Signaling, #5625), rabbit anti-phospho-SAPK/JNK (Thr183/Tyr185) (1:100, Cell Signaling, #9251), rabbit anti-TNFR1 (1:100, Abcam, #19140) or rabbit anti-γH2AX (1:100, Cell Signaling, #9718) overnight at 4 °C. Samples were subsequently stained with AlexaFluor 488-conjugated goat anti-rabbit secondary antibody (1:400) for 1 h and analyzed on a FACS Calibur (Becton Dickinson). Data were analyzed with FlowJo software.
3
Autophagy Modulation in Cancer Cells
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Dabrafenib, 3-methyladenosine (3-MA) (#M9218), and chloroquine (CQ) (#C6628) were purchased from Sigma (St. Louis, MO, USA). The primary antibodies used in this study were rabbit anti-cleaved PARP (#5625, 1:1000, Cell Signaling Technology, Danvers, MA, USA), mouse anti-β-actin (#3700, 1:3000, Cell Signaling Technology, Danvers, MA, USA), mouse anti-LC3I/II (NB600-1384, 1:500, Novus, Littleton, CO, USA), mouse anti-p62 (SC-48389, 1:500, Santa Cruz, CA, USA), and rabbit anti-HMGB1 (ab79823, 1:200, Abcam, Cambridge, MA, USA). Species-specific secondary antibodies were obtained from LI-COR Biosciences.
4
Western Blot Analysis of Apoptotic Markers
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At the end of the incubation periods, islets and INS-1E cells were washed in ice-cold PBS and lysed in RIPA lysis buffer containing 50 mM Tris HCl, pH 8, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% SDS supplemented with Protease- and phosphatase inhibitors (Pierce, Rockford, IL, USA). Protein concentrations were determined with the BCA protein assay (Pierce). Equivalent amounts of protein from each treatment group were run on a NuPAGE 4–12% Bis-Tris gel (Invitrogen) and electrically transferred onto PVDF membranes. After blocking by 2.5% milk (Cell Signaling) and 2.5% BSA, membranes were incubated overnight at 4 °C with rabbit anti-cleaved caspase-3 (#9664), rabbit anti-PARP (#9532), rabbit anti-cleaved PARP (rat specific #9545), rabbit anti-phospho YAP(S127) (#4911), rabbit anti-LATS2 (#5888), rabbit anti-tubulin (#2146), rabbit anti-GAPDH (#2118), rabbit anti-β-actin (#4967) (all Cell Signaling Technology), and rabbit anti-PDX1 (#47267) and rabbit anti-p-MST1 (#79199) (both from Abcam) antibodies, all at a dilution of 1:1000, followed by horseradish-peroxidase-linked anti-rabbit IgG (Jackson). Membrane was developed by using a chemiluminescence assay system (Pierce) and analyzed using DocIT®LS image acquisition 6.6a (UVP BioImaging Systems, Upland, CA, USA). Uncropped and unprocessed scans of all Western blots are available in the Source Data file.
5
Western Blot Analysis of Cell Signaling
6
Prostaglandin Signaling Pathway Modulation
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Oxaliplatin, N-acetyl-L-cysteine (NAC) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich, MO. PGE2, EP receptor selective antagonists and EP4 receptor agonist were purchased from Cayman Chemicals, MI. Antibodies used for immunoblotting and immunofluorescence were as follows: rabbit anti-mPGES-1 (Abnova, Taiwan), rabbit anti-COX-2, rabbit anti-EP1, rabbit anti-EP2, rabbit anti-EP3, rabbit anti-EP4 (Cayman Chemicals), rabbit anti-cleaved PARP, rabbit anti-phospho-Akt, rabbit anti-Bcl2, rabbit anti-Bax (Cell Signaling Technology, MA), mouse anti-β actin (Sigma). Mouse anti-15-PGDH was a generous gift from Drs. Sanford D. Markowitz and Stephen Fink at Case Western Reserve University.
7
Western Blot Analysis of Lipogenesis Proteins
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Immunoblots were performed as described previously (5 (link)). Briefly, cell pellets was suspended in RIPA lysis buffer (5 (link)). Protein concentration was determined using bicinchoninic acid assay (Pierce). Equal amount of total protein was loaded and separated by SDS-PAGE and transferred onto polyvinylidene fluoride membrane (Millipore). The membranes were blocked with 4% BSA/Tris-buffered saline with 0.1% Tween 20 detergent (TBS-T) and then probed with corresponding antibodies. The following antibodies were used: mouse anti-β-actin (Sigma, #A5316, RRID:AB_476743), rabbit anti-SCD1 (Abcam, #ab236868, RRID: AB_2928123), rabbit anti-CARM1 (Cell Signaling Technology, #3379S, RRID:AB_2068433), rabbit anti-FASN (Cell Signaling Technology, #3189, RRID:AB_2100798), rabbit anti-ACC1 (Cell Signaling Technology, #4190, RRID:AB_10547752), rabbit anti-cleaved PARP (Cell Signaling Technology, #5625S, RRID:AB_10699459), and rabbit anti-Lamin A/C (Cell Signaling Technology, #2032S, RRID:AB_2136278).
8
Antibody Profiling for Apoptosis Signaling
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Antibodies used for immunoblotting analyses in this study are listed below: rabbit anti-MCL-1 (1:500, ab32087), rabbit anti-BAX (1:1000, ab32503) were obtained from Abcam; mouse anti-BCL-2 (1:1000, #15071), rabbit anti-BIM (1:1000, #2933), rabbit anti-BCL-XL (1:1000, #2762), rabbit anti-BAK (1:1000, #12105), rabbit anti-AKT (1:1000, #4691), rabbit anti-p-AKT (1:1000, #4060), rabbit anti-mTOR (1:1000, #2983), rabbit anti-p-mTOR (1:1000, #5536), rabbit anti-p-4EBP1 (1:1000, #2855), rabbit anti-4EBP1 (1:1500, #9644), rabbit anti-FOXO3a (1:1000, #12829), rabbit anti-p-FOXO3a (1:1000, #9466) rabbit anti-cleaved PARP (1:1000, #5625) and rabbit anti-cleaved Caspase-3 (1:1000, #9664) were purchased from Cell Signaling Technology; mouse anti-β-actin (1:1000, sc-47778), mouse anti-GAPDH (1:1000, sc-32233) and rabbit anti-PUMA (1:1000, sc-28226) were from Santa Cruz Biotechnology; rabbit anti-p-4EBP1 (1:500, NB100-81769, used in Fig. 1b ) and mouse anti-4EBP1 (1:1000, NBP1-47366, used in Fig. 1b ) were obtained from Novus Biologicals. The compounds of BH3 mimetics (ABT263 and ABT199) and mTOR inhibitors (BEZ235, AZD8055, and Temsirolimus) were from AbMole Bioscience.
9
Signaling Pathway Analysis in Cell Lines
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BCA (purity, 95.8%) was purchased from the Institute for Korea Traditional Medical Industry (Daegu, Korea) and dissolved in DMSO (Sigma-Aldrich, St. Louis, MO, USA). A BCA stock solution of 40 mM was stored at -80°C. Mouse anti-β-actin (1 : 5000 dilution), rabbit anti-p-AKT (1 : 1000 dilution), rabbit anti-AKT (1 : 1000 dilution), rabbit anti-p-p53 (Ser15) (1 : 1000 dilution), rabbit anti-p-p53 (Ser20) (1 : 1000 dilution), rabbit anti-p-p53 (Ser46) (1 : 1000 dilution), and rabbit anti-MDM2 (1 : 1000 dilution) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit anti-p21 (1 : 1000 dilution), rabbit anti-p27 (1 : 1000 dilution), rabbit anti-p53 (1 : 1000 dilution), rabbit anti-FOXO3 (1 : 1000 dilution), rabbit anti-Bcl-2 (1 : 1000 dilution), rabbit anti-Bcl-xL (1 : 1000 dilution), rabbit anti-Bax (1 : 1000 dilution), rabbit anti-cleaved caspase-3 (1 : 1000 dilution), rabbit anti-caspase-3 (1 : 1000 dilution), rabbit anti-cleaved caspase-7 (1 : 1000 dilution), rabbit anti-caspase-7 (1 : 1000 dilution), rabbit anti-cleaved caspase-9 (1 : 1000 dilution), rabbit anti-caspase-9 (1 : 1000 dilution), rabbit anti-cleaved PARP (1 : 1000 dilution), and rabbit anti-PARP (1 : 1000 dilution) were purchased from Cell Signaling Technology (Danvers, MA, USA).
10
Immunoblot Analysis of mTOR Signaling
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The antibodies used in this study include the following: rabbit anti-phosphor-mTOR, rabbit anti-mTOR, mouse anti-Phospho-p70 S6 Kinase, rabbit anti-p70 S6 kinase, rabbit anti-CD44, rabbit anti-cleaved PARP and rabbit anti-cleaved caspase-3 were all obtained from Cell Signaling Technology (Danvers, MA, USA). Mouse-anti-p32 (gC1qR) antibody was obtained from Abcam (Cambridge, MA, USA). Rabbit anti-Akt and mouse anti-Phospho-Akt antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Goat anti-mouse and anti-rabbit IgG-horseradish peroxidase-conjugates were from Pierce (Rockford, IL, USA). Mouse monoclonal anti-actin was a gift from Dr. José Manuel Hernandez-Hernández (CINVESTAV-IPN). Temsirolimus (CCI-779) and 5-fluorouracil (5-FU) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
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