Cleaved caspase 3

Manufactured by New England Biolabs

Sourced in United Kingdom, United States

Cleaved caspase-3 is an enzyme that plays a key role in the process of apoptosis, or programmed cell death. It is produced when the inactive precursor caspase-3 is cleaved and activated. Cleaved caspase-3 functions as a central executioner in the apoptotic pathway, leading to the breakdown of cellular structures and the death of the cell.

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Lab products found in correlation

X ray film, by Fujifilm (2 mentions) Enhanced chemiluminescence, by GE Healthcare (1 mentions) Op46 100ug, by Merck Group (1 mentions) Phospho p53 ser15, by Cell Signaling Technology (1 mentions) Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions) Ncl l p53 do7, by Leica (1 mentions) Hybond c extra membrane, by GE Healthcare (1 mentions) Mdr 1, by Santa Cruz Biotechnology (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) Mini protean tgx precast gel, by Bio-Rad (1 mentions) Phospho akt ser473, by Santa Cruz Biotechnology (1 mentions)

3 protocols using cleaved caspase 3

Western Blotting of Cell Signaling Proteins

Cited 1 time

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Western blotting was carried out as described in (31 (link)). Antibodies used were MDM2 (Ab-1) 1:300 (#: OP46-100UG, Merck Millipore), MDMX (#: A300287A-2 Bethyl laboratories), WIP1 (F-10) 1:200 (#: sc-376257, Santa Cruz Biotechnology), p53 1:500 (#: NCL-L-p53-DO7, Leica Microsystems Ltd.), phospho-p53^Ser-15 1:1000 (#: 9284 Cell Signalling), p21^WAF1 1:100 (#: OP64, Calbiochem), BAX 1:1000 (#: 2772S, Cell Signalling), cleaved caspase-3 1:1000 (#: 9664S, New England Biolabs Ltd.), actin 1:3000 (#: A4700, Sigma-Aldrich). Secondary goat anti-mouse/rabbit HRP-conjugated antibodies (#: P0447/P0448, Dako) were used at 1:1000. All antibodies were diluted in 5% milk/1 × TBS-tween (w/v). Proteins were visualised using enhanced chemiluminescence (GE Life Sciences) and X-ray film (Fujifilm). Densitometry was carried out using ImageJ software.

Esfandiari A., Hawthorne T.A., Nakjang S, & Lunec J. (2016). Chemical inhibition of wild-type p53 induced phosphatase 1 (WIP1/PPM1D) by GSK2830371 potentiates the sensitivity to MDM2 inhibitors in a p53-dependent manner. Molecular cancer therapeutics, 15(3), 379-391.

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Western Blot Analysis of Protein Biomarkers

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Whole-cell lysates were harvested as previously described (Tweddle et al, 2001b (link)). Proteins were separated using 4–20% Mini-PROTEAN TGX Precast Gels (Bio-Rad Laboratories Ltd, Hemel Hempstead, UK) and transferred onto Hybond-C Extra membrane (GE Life Sciences, Little Chalfont, UK). Primary antibodies used were MDR-1 1 : 200 (Cat No.: sc-13131, Santa Cruz Biotechnology Inc., Dallas, TX, USA), p53 1 : 1000 (Cat No.: NCL-L-p53-DO7, Leica Microsystems Ltd, Newcastle upon Tyne, UK), MYCN 1 : 100 (Cat No.: OP13, Merck KGaA), cleaved caspase-3 1 : 1000 (Cat No.: 9664S, New England Biolabs Ltd, Hitchin, UK), actin 1 : 500 (Cat No.: A4700, Sigma-Aldrich) and GAPDH 1 : 500 (Cat No.: sc-25778, Santa Cruz Biotechnology Inc.). Secondary goat anti-mouse/rabbit HRP-conjugated antibodies (Cat No.: P0447/P0448, Dako, Glostrup, Denmark) were used at 1 : 1000. All antibodies were diluted in 5% milk/1 × TBS-tween (w/v). Protein detection was performed using enhanced chemiluminescence (GE Life Sciences) and X-ray film (Fujifilm, Bedford, UK).

Chen L., Zhao Y., Halliday G.C., Berry P., Rousseau R.F., Middleton S.A., Nichols G.L., Del Bello F., Piergentili A., Newell D.R., Lunec J, & Tweddle D.A. (2014). Structurally diverse MDM2–p53 antagonists act as modulators of MDR-1 function in neuroblastoma. British Journal of Cancer, 111(4), 716-725.

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Proanthocyanidin-Mediated Cellular Signaling

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GSE was donated by the INDENA Company (Milan, Italy) and used throughout the study. The GSE contains approximately 93.8% proanthocyanidins, with monomers (7.6%) dimers (8.3%), and oligomers (77.9%). Proanthocyanidin B2 (GSPB2) was obtained from Sigma (St. Louis MO). Antibodies to phospho-ERK1/2 (Thr202/Tyr204), AKT, phospho-BAD (phospho-Ser-136), BAD, cleaved-caspase-3, and total caspase-3 were purchased from New England Biolabs Inc. (Beverly, MA, USA) and antibodies to ERK2 (C14), cyclin D2 (C17), p21, p27, and phospho-AKT (Ser 473) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Monoclonal antibodies to vinculin were purchased from Sigma. Rabbit polyclonal antibodies against StAR were generously provided by Dale Buchanan Hales (University of Illinois, Chicago, IL, USA). All antibodies were used at 1/1000 dilution in Western blotting.

Barbe A., Ramé C., Mellouk N., Estienne A., Bongrani A., Brossaud A., Riva A., Guérif F., Froment P, & Dupont J. (2019). Effects of Grape Seed Extract and Proanthocyanidin B2 on In Vitro Proliferation, Viability, Steroidogenesis, Oxidative Stress, and Cell Signaling in Human Granulosa Cells. International Journal of Molecular Sciences, 20(17), 4215.

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