Antibodies for caspase 3

Manufactured by Cell Signaling Technology

Sourced in United States

Antibodies for caspase-3 are laboratory reagents used to detect and quantify the presence of caspase-3, a key enzyme involved in the apoptosis (programmed cell death) process. These antibodies can be used in various analytical techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the role of caspase-3 in cellular processes.

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Caspase-3 (2138 citations) Antibodies for cleaved caspase-3 (8 citations) Primary antibodies for caspase-3 (5 citations) Antibodies specific for Cleaved Caspase-3, (3 citations) Caspases (2 citations) Antibodies specific for caspase-3 (2 citations)

5 protocols using antibodies for caspase 3

Western Blotting Protein Expression Analysis

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The cells were harvested and lysed in RIPA buffer (50mM Tris-HCl pH 7.5, 150mM NaCl, 1% IPEGAL, 0.5% deoxycholate, 5mM EDTA) containing a protease inhibitor cocktail (Roche, Mannheim, Germany). For protein immunoblot analysis, polypeptides in whole cell lysates were resolved by SDS-PAGE and transferred to nitrocellulose membranes. The images were acquired using Odyssey (Licor bioscience, Lincoln, NE). The antibodies for NOX1 (Cat #:7921), NOX2 (Cat #:7923), NOX3 (Cat #:7925) and NOX4 (Cat #:7927) were from ProSci (Poway, CA). The antibodies for Caspase 3 (Cat #:9668), Cyclin D1 (Cat #: 2978), S6 Ribosomal Protein antibody (5G10), phospho S6 ribosomal protein [pSer235/236] (D57.2.2E) were from Cell Signaling Technology (Beverly, MA). The antibodies for Akt1 antibody (Y89), phospho-Akt [pSer473] antibody (EP2109Y) were from Abcam (Cambridge, MA). The antibody for NOX5 from Boster Biological Technology (Pleasanton, CA), the antibody for DUOX1 from GeneTex (Irvine, CA), the antibody for DUOX2 from Novus Biologicals (Littleton, CO), and the antibody for p22phox from Santa Cruz Biotechnology (San Diego, CA).

Jafari N., Kim H., Park R., Li L., Jang M., Morris A.J., Park J, & Huang C. (2017). CRISPR-Cas9 Mediated NOX4 Knockout Inhibits Cell Proliferation and Invasion in HeLa Cells. PLoS ONE, 12(1), e0170327.

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Liver Protein Fractions Analysis

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Total, cytosolic, mitochondrial, and microsomal liver protein fractions were separated by electrophoresis using polyacrylamide gels and transferred to PVDF membranes. Membranes were incubated overnight (4 °C) with antibodies for caspase-3 (Cat# 14220, Cell Signaling Technology, Danvers, MA), Drp-1 (Cat# 8570), cytochrome c (Cat# sc-13156, Santa Cruz Biotechnology, Dallas, TX), Sirt3 (Cat# sc-365175), Tlr9 (Cat# sc-52966), β-actin (Cat# sc-47778), Gapdh (Cat# sc-32233) or Cox-IV (Cat# sc-517553), followed by IRDye® donkey anti-rabbit or goat anti-mouse IgG (H+L) 680 RD and goat anti-mouse IRDye® 800 CW (LI-COR Biosciences, Lincoln, NE) at room temperature for 1 hour. Blots were imaged using LI-COR Odyssey® Clx scanner. Protein band density was determined using Image J. Bands were normalized to β-actin (whole lysate), Gapdh (cytosol, microsomal), or Cox-IV (mitochondria) protein levels, and expressed as fold-change vs. sham (n = 3–5/group).

Hou J., Tolbert E., Birkenbach M, & Ghonem N.S. (2021). Treprostinil alleviates hepatic mitochondrial injury during rat renal ischemia-reperfusion injury. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 143, 112172.

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NR4A2 Regulation of Autophagy and Apoptosis

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Dulbecco's modified Eagle's medium (DMEM) was purchased from Gibco (Grand Island, NY, USA). NR4A2 and negative control siRNA were purchased from Ribbio (Guangzhou, China). 3MA, Bafilomycin A1 and NH₄Cl were purchased from Sangon Biotech (Shanghai, China). Lipofectamine 2000 was purchased from Invitrogen (Carlsbad, CA). NR4A2 antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies for caspase3, PARP, LC3, p53 and Bax were from Cell Signaling Technology (Beverly, MA, USA).

Liu H., Liu P., Shi X., Yin D, & Zhao J. (2018). NR4A2 protects cardiomyocytes against myocardial infarction injury by promoting autophagy. Cell Death Discovery, 4, 27.

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Quantitative Western Blot Analysis

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Antibodies for caspase-3, caspase-9, phospho-p38, phospho-JNK, phospho-ASK1 and ASK1, and β-actin were purchased from Cell Signaling Technology (Danvers, MA). Antibodies for JNK, iNOS and COX-2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). The protein content of liver homogenates was measured using a bicinchoninic acid (BCA) kit (Thermo Scientific, Rockford, IL). Protein aliquots (10-40 μg) were electrophoresed on 4%–20% gradient SDS-PAGE gels (Life Technologies, Grand Island, NY) and then transferred to PVDF membranes (Millipore, Billerica, MA). The blocked membranes were incubated with primary antibody in tris-buffered saline tween 20 (TBST) containing 5% nonfat milk or BSA at 4°C overnight, followed by incubation with the appropriate secondary antibodies (Cell Signaling Technology) according to the manufacturer's instructions. Membranes were developed with super-signal west femto maximum sensitivity substrate (Thermo Scientific). Protein loading was routinely confirmed with an antibody against β-actin. Densitometric analysis was conducted using Image Lab Software (Bio-Rad, Hercules, CA). Quantitative analysis was performed by calculating the densitometry ratios versus β-actin.

Zhao Y., Zhou P., Liu B., Bambakidis T., Mazitschek R., Alam H.B, & Li Y. (2014). Protective Effect of SAHA against LPS-induced Liver Damage in Rodents. The Journal of surgical research, 194(2), 544-550.

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Apoptosis Induction Assay

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EGCG, EGC, ECG, EC, C, GC, GCG, CG, 3-(4,5-dimethylthiaxol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), propidium iodide (PI), and antibodies for β-actin were purchased from Sigma Chemical Co (St. Louis, MO, USA). Dimethyl sulfoxide (DMSO) and sodium dodecyl sulfate (SDS) were purchased from Merck Co (Darmstadt, Germany). Antibodies for caspase-3, caspase-9, poly(ADP-ribose) polymerase (PARP), p21, and p53 were purchased from Cell Signaling Technology (Beverly, MA, USA).

Wang C.C., Ho C.T., Lee S.C, & Way T.D. (2015). Isolation of eugenyl β-primeveroside from Camellia sasanqua and its anticancer activity in PC3 prostate cancer cells. Journal of Food and Drug Analysis, 24(1), 105-111.

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