Caspase 3 clone 8g10

Manufactured by Cell Signaling Technology

Sourced in United States

Caspase-3 (clone 8G10) is a laboratory antibody product offered by Cell Signaling Technology. It recognizes the Caspase-3 protein, which is a key enzyme involved in the execution phase of cell apoptosis (programmed cell death).

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Lab products found in correlation

Nigericin, by Cayman Chemical (1 mentions) Vildagliptin, by Cayman Chemical (1 mentions) Saxagliptin, by LGC (1 mentions) Sitagliptin, by Merck Group (1 mentions) Human il 1β, by R&D Systems (1 mentions) Lipopolysaccharides (lps), by Santa Cruz Biotechnology (1 mentions) Etoposide, by Enzo Life Sciences (1 mentions) Ala pro afc, by Bachem (1 mentions) Human caspase 1, by Cell Signaling Technology (1 mentions) Gsdmd nbp2 33422, by Novus Biologicals (1 mentions) Gapdh clone 14c10, by Cell Signaling Technology (1 mentions) Ac yvad cmk, by Cayman Chemical (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Z vad fmk, by Enzo Life Sciences (1 mentions) Il 1α, by R&D Systems (1 mentions) Il 18, by R&D Systems (1 mentions) Rapamycin, by Merck Group (1 mentions) Abt 199, by Selleck Chemicals (1 mentions) Phosphor p70s6k thr389, by Cell Signaling Technology (1 mentions) Noxa sc 56169, by Santa Cruz Biotechnology (1 mentions)

4 protocols using caspase 3 clone 8g10

Caspase Activation and Pyroptosis Assays

Cited 1 time

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LPS was purchased from Santa Cruz Biotechnology, nigericin, and vildagliptin and Ac-YVAD-CMK from the Cayman Chemical Company, PMA and sitagliptin from Sigma, Ala-Pro-AFC from Bachem, saxagliptin from Toronto Research Chemicals, and Z-VAD-FMK and etoposide from Enzo Life Sciences. Val-boroPro^{45 (link)}, 1G244^{24 (link)}, FP-biotin^{15 (link)}, L-allo-Ile-isoindoline^{14 (link)}, and L-allo-Ile-thiazolidine^{14 (link)} were synthesized according to previously published protocols. For cell culture experiments, Val-boroPro was resuspended in DMSO containing 0.1% TFA to prevent compound cyclization. Antibodies used include: human caspase-1 (#2225, Cell Signaling Technology), mouse caspase-1 (clone Casper-1, Adipogen), caspase-3 (clone 8G10, Cell Signaling Technology), human caspase-4 (clone 4B9, Santa Cruz), human caspase-5 (clone D3G4W, Cell Signaling Technology), caspase-7 (clone D2Q3L, Cell Signaling Technology), human IL-1β (Clone 2805, R&D Systems), mouse IL-1β (clone D4T2D, Cell Signaling Technology), IL-1α (#AF-200, R&D Systems), IL-18 (#AF2548, R&D Systems), GAPDH (clone 14C10, Cell Signaling Technology), DPP7 (Clone 398024, R&D Systems), DPP8 (ab42076, Abcam), DPP9 (ab42080, Abcam), PARP (#9542, Cell Signaling Technology), GSDMD (NBP2-33422, Novus Biologicals), DPP4 (#11D7, GeneTex), FAP (ABT11, Millipore), and SCPEP1 (SAB2700267, Sigma).

Okondo M.C., Johnson D.C., Sridharan R., Go E.B., Chui A.J., Wang M.S., Poplawski S.E., Wu W., Liu Y., Lai J.H., Sanford D.G., Arciprete M.O., Golub T.R., Bachovchin W.W, & Bachovchin D.A. (2016). DPP8/9 inhibition induces pro-caspase-1-dependent monocyte and macrophage pyroptosis. Nature chemical biology, 13(1), 46-53.

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Apoptosis Pathway Regulation Protocol

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2-DG, PA, SP600125, glucose, rapamycin, the proteasome inhibitor MG132, Annexin V and PI were obtained from Sigma (St. Louis, MO, USA). ABT-199 was from Selleck Chemicals (Houston, TX, USA). myc (clone 9E10, M4439) and actin (clone AC-74, A5316) antibodies were also from Sigma. phospho-Akt (Ser 473) (clone 587F11, #4051), Akt (#9272), phosphor-mTOR (Ser2448)(#5536), mTOR (#2972), phosphor-p70S6K (Thr389)) (#9234), p70S6K (#9202), phospho-JNK (Thr 183/Tyr 185) (#9251), JNK (#9252), caspase-3 (clone 8G10, #9665), and Mcl-1 (#4572) antibodies were purchased from Cell Signaling (Beverly, MA). Bcl-2 (sc-7382), Bcl-xL (sc-8392), phosphor-Bcl-xL (Ser 62) (sc-101644), BimEL (sc-27982), p62(sc-55603), Noxa (sc-56169) and Bid (sc-56025) antibodies were from Santa Cruz (Santa Cruz, CA).

Zhang Y., Li X., Tan S., Liu X., Zhao X., Yuan Z, & Nie C. (2016). Mcl-1 expression and JNK activation induces a threshold for apoptosis in Bcl-xL-overexpressing hematopoietic cells. Oncotarget, 8(7), 11042-11052.

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Western Blot Analysis of Proteins

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Proteins were isolated from tissues, resolved by SDS-PAGE then transferred to PVDF membrane as previously described.⁸ Membranes were immunoblotted with the primary antibodies specific to: caspase-3 (clone 8G10, #9665), cleaved caspase-3 (#9664) and p53 (clone 1C12, #2524; Cell Signalling Technology, Beverly, MA, USA); UCP1 (#ab10983; Abcam, Cambridge, MA, USA); and β-actin clone AC-15 (#A5441; Sigma-Aldrich, St. Louis, MO, USA). Densitrometric analysis was performed using Image J software (NIH, Bethesda, MD, USA).

Wilson C.H., Nikolic A., Kentish S.J., Keller M., Hatzinikolas G., Dorstyn L., Page A.J, & Kumar S. (2017). Caspase-2 deficiency enhances whole-body carbohydrate utilisation and prevents high-fat diet-induced obesity. Cell Death & Disease, 8(10), e3136-.

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Inhibition of DNA Repair Pathways

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Cells were plated in 10-cm dishes and allowed to attach overnight. AZD6738 and KU-0060648 were added at the indicated concentrations 1 h before irradiation. Medium and cells were harvested in PBS-containing 1 mM Na₃VO₄ and 1 mM NaF. Cells were pelleted before lysis in 50 mM Tris–HCl pH 7.5, 150 mM NaCl, 1% NP-40, 0.5% deoxycholate, and 0.1% SDS. Lysis supernatants were quantified by BCA assay from Pierce (Leicestershire, UK). Total protein lysate was separated by reducing SDS-PAGE, transferred to Midi Nitrocellulose membranes using a Transblot Turbo (Bio-Rad, Watford, UK), and blocked with TBS blocking buffer (Li-Cor Biotechnology, Cambridge, UK). Membranes were probed with antibodies specific for GAPDH clone 10C10, CHK1 clone 2G1D5, phospho-CHK1 (S345) clone 133D3, DNA-PK code 4602, γH2Ax (S139) clone 20E3 and Caspase 3 clone 8G10 from Cell Signaling (MA, USA); phospho-DNA-PK from Abcam ab18192 (Cambridge, UK); PARP clone F-2 was purchased from Santa Cruz (TX, USA).

Hafsi H., Dillon M.T., Barker H.E., Kyula J.N., Schick U., Paget J.T., Smith H.G., Pedersen M., McLaughlin M, & Harrington K.J. (2018). Combined ATR and DNA-PK Inhibition Radiosensitizes Tumor Cells Independently of Their p53 Status. Frontiers in Oncology, 8, 245.

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