Caspase 3

Manufactured by Santa Cruz Biotechnology

Sourced in United States, China, United Kingdom, Canada, Germany, France

Caspase-3 is a key enzyme involved in the execution phase of cell apoptosis. It is a protease that cleaves specific substrates, leading to the characteristic morphological and biochemical changes associated with programmed cell death.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Mouse anti-caspase-3 antibody (14 citations) Rabbit anti-cleaved caspase-3 (9 citations) Anti-cleaved caspase-3 antibody (4 citations) Caspase-3 (sc-7148) (4 citations) Caspase-3 primary antibody (4 citations) Caspase-3 p17 (3 citations) Caspase-3 (H-277), (3 citations) Full-length caspase-3 (2 citations) Anti-caspase-3 p30/17 (2 citations) Caspase-3 Apoptosis Detection Kit (2 citations)

428 protocols using caspase 3

Protein Expression Analysis by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot assay was conducted as previously described [17 (link)]. The proteins from the cell were lysed in RIPA buffer (pH = 7.4) comprised of protease inhibitors cocktail (10 μg/ml, Cat# 5872S, Cell Signaling Technology, MA, U.S.A.). The contents were centrifuged at 12,000 g at 4°C for 20 min, and the concentration of protein in the supernatants was determined using Bradford reagent (BioRad) with bovine serum albumin (BSA, Sigma Aldrich, St. Louis, MO, U.S.A.) as the standard. The protein samples were separated by electrophoresis on SDS-PAGE 10% or 12.5% gels. After blocked in 3% BSA, the membrane was incubated with primary antibodies, GAPDH (Cat# 2118 s, Cell Signaling Technology), caspase-1 (Cat# sc-56036, Santa Cruz Biotechnology, U.S.A.), GSDMD (Cat# 93709 s, Cell Signaling Technology), JNK1/2/3 (Cat# YT2440, Immunoway, TX, U.S.A.), caspase-3 (Cat# sc-7148, Santa Cruz Biotechnology), caspase-9 (Cat# 9502, CST), caspase-4 (Cat# ab238124, Abcam, Cambridge, United Kingdom), cleaved caspase-3 (Cat# 9661 s, Cell Signaling Technology), and GSDME (Cat# ab215191, Abcam) as needed. For secondary antibodies, antibodies to mouse (Cat# 7076, Cell Signaling Technology) and rabbit (Cat# 7074, Cell Signaling Technology) were used. To visualize protein bands, a chemiluminescence (ECL) system (Cat# WBLUF0500, Millipore, MA, U.S.A.) was used.

Zhang F., Liu Q., Ganesan K., Kewu Z., Shen J., Gang F., Luo X, & Chen J. (2021). The Antitriple Negative Breast cancer Efficacy of Spatholobus suberectus Dunn on ROS-Induced Noncanonical Inflammasome Pyroptotic Pathway. Oxidative Medicine and Cellular Longevity, 2021, 5187569.

+ Open protocol

+ Expand

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein extraction was carried out as follows: cardiac tissues were isolated, snap‐frozen and extracted in lysis buffer, and cardiomyocytes were harvested at the indicated time and extracted in lysis buffer. The lysis buffer was complemented with phenylmethanesulfonyl fluoride and phosphatase inhibitor. BCA Protein Assay Kit (Thermo Scientific, Rockford, IL, USA) was used to quantify total protein concentration. Protein samples were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred onto a polyvinylidene fluoride membrane. The membrane was blocked with 5% skim milk and incubated at 4°C overnight with the following primary antibodies: phospho‐adenosine monophosphate protein kinase (AMPK)‐T172, AMPK, p‐Raptor‐S792, Raptor, p‐mTOR‐S2448, mTOR, p‐p70S6K‐T389, p70S6K, β‐actin (1:1000; Cell Signaling Technology, Danvers, MA, USA) and caspase‐3 (1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA). The membrane was then incubated with the appropriate horseradish peroxidase‐conjugated secondary antibodies. The blots were developed using an enhanced chemiluminescence system.

Wang D., Jiang L., Feng B., He N., Zhang Y, & Ye H. (2019). Protective effects of glucagon‐like peptide‐1 on cardiac remodeling by inhibiting oxidative stress through mammalian target of rapamycin complex 1/p70 ribosomal protein S6 kinase pathway in diabetes mellitus. Journal of Diabetes Investigation, 11(1), 39-51.

+ Open protocol

+ Expand

Diosgenin-Induced Apoptosis Mechanisms

Check if the same lab product or an alternative is used in the 5 most similar protocols

Diosgenin, RIPA buffer was obtained from Sigma Aldrich (St. Louis, MO, USA). RPMI-1640, DMEM medium, fetal bovine serum (FBS), trypsin, penicillin, and streptomycin were purchased from Gibco (Rockville, MD, USA). PI (propidium iodide), Annexin V-FITC/PI apoptosis detection kit, z-VAD-fmk, MTS, Hoechst 33258, and AO/EB (acridine orange/ethidium bromide) were purchased from Abcam (Cambridge, United Kingdom). 5-Ethynyl-2-deoxyuridine (EdU) was obtained from RiboBio (Guangzhou, China). The primary antibodies against GAPDH, Bax, Bcl-2, Caspase- 3, p21, PARP-1, cytochrome c (CYT C), COX IV, β-catenin and GSK3β were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Other antibodies were from Abcam. Hematoxylin–Eosin Staining Kit was from Solarbio (Beijing, China). The EliVision kit was from Maixin Biotech (Fuzhou, China).

Mao X.M., Zhou P., Li S.Y., Zhang X.Y., Shen J.X., Chen Q.X., Zhuang J.X, & Shen D.Y. (2019). Diosgenin Suppresses Cholangiocarcinoma Cells Via Inducing Cell Cycle Arrest And Mitochondria-Mediated Apoptosis. OncoTargets and therapy, 12, 9093-9104.

+ Open protocol

+ Expand

Western Blot Analysis of Apoptosis Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Following cell lysis by ice-cold radioimmunoprecipitation assay buffer, protein concentrations were determined. Equal quantities (30 µg) of protein were separated using 10% SDS-polyacrylamide gels, transferred to a polyvinylidene difluoride (PVDF) membrane and blocked with 5% bovine serum albumin (BSA). PVDF membranes were incubated overnight at 4°C with the following antibodies: CIP2A (dilution 1:500; cat. no. sc-80662; Santa Cruz Biotechnology, Inc., Dallas, CA, USA), AKT (dilution 1:1,000; cat. no. 9272S; Cell Signaling Technology, Inc., Danvers, MA, USA), phosphorylated (p)-AKT (dilution 1:1,000; cat. no. 4060S; Cell Signaling Technology), B cell lymphoma-2 (Bcl-2; dilution 1:1,000; cat. no. sc-492; Santa Cruz Biotechnology, Inc.), caspase-3 (dilution 1:1,000; cat. no. 9662S; Cell Signaling Technology), poly(ADP-Ribose) polymerase (PARP; dilution 1:1,000; cat. no. 5625T; Cell Signaling Technology), γ-H2AX (dilution 1:1,000; cat. no. ab2893; Abcam, Cambridge, UK) and β-actin (dilution 1:1,000; cat. no. sc-47778; Santa Cruz Biotechnology, Inc.). After being washed, the membranes were then incubated with horseradish peroxidase-conjugated secondary antibodies (anti-mouse IgG, dilution 1:1,000; cat. no. 7076S; Cell Signaling Technology; anti-rabbit IgG, dilution 1:1,000; cat. no. 7074S; Cell Signaling Technology) followed by enhanced chemiluminescence detection.

Gao F., Wang X., Chen S., Xu T., Wang X., Shen Y., Dong F., Zhong S, & Shen Z. (2018). CIP2A depletion potentiates the chemosensitivity of cisplatin by inducing increased apoptosis in bladder cancer cells. Oncology Reports, 40(5), 2445-2454.

+ Open protocol

+ Expand

Wogonin's Anti-Cancer Potential Explored

Check if the same lab product or an alternative is used in the 5 most similar protocols

A2780 cells were purchased from Sigma-Aldrich Co. (St Louis, MO) and were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS, Gibco). Cells were tested with a Cell Culture Contamination Detection Kit (Thermo Fisher Scientific) and results appeared negative for mycoplasma contamination. Wogonin, with a chemical structure shown in Figure 1(a), was purchased from Aokebio (Beijing, China). Methylpiperidinopyrazole (MPP) was purchased from Apexbio. The antibodies were from Abcam (Akt, β-actin), Proteintech (caspase-3, cleaved-caspase-3, cyclin D1, CDK4, and CDK6), Santa Cruz Biotechnology (ER-α), and Cell Signaling Technology (Bcl-2, Bax, VEGF, and p53).

Ruibin J., Bo J., Danying W., Chihong Z., Jianguo F, & Linhui G. (2017). Therapy Effects of Wogonin on Ovarian Cancer Cells. BioMed Research International, 2017, 9381513.

+ Open protocol

+ Expand

Western Blotting and Immunofluorescence Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols

For Western blotting, whole cell lysates were prepared by lysing cell pellets directly in sodium dodecyl (SDS)-polyacrylamide gel electrophoresis buffer. For immunofluorescence with the Mcm3 antibody, HeLa cells were pre-extracted with phosphate-buffered saline (PBS) containing 0.5% (v/v) Triton X-100, and fixed in 4% paraformaldehyde solution (WAKO) for immunostaining. Images were acquired by Keyence BZ-8100 and nuclear staining intensity was measured by Dynamic Cell count software (Keyence). The following primary antibodies were used: Cdt1[6 (link)], Cdt2[50 (link)], Mcm2 (lab stock), Mcm3 (ab4460, Abcam, Cambridge, UK), Mcm4 (lab stock), Mcm6 (Santa Cruz Biotechnology, Dallas, TX), Cyclin A (mouse, Ab-6, Neomarkers; rabbit, H-432, Santa Cruz Biotechnology), Cyclin B (H433, Santa Cruz Biotechnology), PCNA (PC10, Santa Cruz Biotechnology), caspase-3 (96625, Santa Cruz Biotechnology), RCC1 (lab stock), Chk1 pS296 (Cell Signaling Technology, Danvers, MA), Chk2 pT68 (Cell Signaling Technology), Cdk2 pT160 (Cell Signaling Technology), and CPD (Cosmo Bio, Tokyo, Japan). Protein levels were analyzed by ImageJ software.

Morino M., Nukina K., Sakaguchi H., Maeda T., Takahara M., Shiomi Y, & Nishitani H. (2015). Mitotic UV Irradiation Induces a DNA Replication-Licensing Defect that Potentiates G1 Arrest Response. PLoS ONE, 10(3), e0120553.

+ Open protocol

+ Expand

Evaluating ECFC-Derived Vascular Formation

Check if the same lab product or an alternative is used in the 5 most similar protocols

After 3 and 28 days following ECFC transplantation, the ischemic thigh areas were removed and fixed with 4 % paraformaldehyde (Affymetrix, Santa Clara, CA, USA). Each tissue sample was embedded in paraffin. For histological analysis, samples were stained with hematoxylin and eosin (H&E). Immunofluorescence staining was performed using primary antibodies against mouse-specific (to confirm mouse vasculature) or human-specific (to confirm human ECFC-derived vessel formation) CD31, α-smooth muscle actin (α-SMA), human VEGF, phospho-STAT3, caspase-3, PCNA, Ki67 (Santa Cruz), human-specific CD31 (Novus Biologicals, Colorado, USA), and human nuclear antigen (HNA; Millipore) and secondary antibodies Alexa-488 and Alexa-594 (Life Technologies, Carlsbad, CA, USA). Nuclei were stained with 4′,6-diamidino-2-phenylindole phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA). Immunostained slides were imaged by confocal microscopy (Olympus, Japan).

Lee S.H., Lee J.H., Han Y.S., Ryu J.M., Yoon Y.M, & Han H.J. (2015). Hypoxia accelerates vascular repair of endothelial colony-forming cells on ischemic injury via STAT3-BCL3 axis. Stem Cell Research & Therapy, 6(1), 139.

+ Open protocol

+ Expand

Lung Cancer Cell Line Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

NSCLC cell lines A549, HCC4006 and Calu-1 (three human lung adenocarcinoma cell lines), and SK-MES-1 and KLN205 (two human lung squamous carcinoma cell lines) were obtained from the Shanghai Institute for Biological Science (China). All cell lines were purchased between 2012 and 2015 and authenticated based on growth rate, morphology, and viability and were frequently confirmed to be mycoplasma free. The cells were cultured in RPMT 1640 media supplemented with 10% heat-inactivated fetal calf serum (FBS), L-glutamine and 100 U/ml penicillin, and 100 μg/ml streptomycin sulfate. The cells were maintained in a humidified incubator in 5% CO₂ at 37 °C.
Antibodies to various antigens were as follows: CHK1, PCHK1 (S296), PCHK1 (S317), PCHK1 (S345), CHK2, PCHK2 (T68), PCHK2 (S516), Cdc25A, PCdc25C (S216), H2AX, γH2AX (S139), PH3 (S10histone), ATM, PATM (S1981), DNA-PKCs, PDNA-PKCs (S2056), and GAPDH were from Cell Signaling. FANCL, FANCD2, BRCA2, PARP1, RAD51, caspase-3, cleaved caspanse-3, PARP and cleaved PARP from Santa Cruz. Gemcitabine was from Hanson Pharmaceutic (China), Cisplatin from Yangtze River Pharmaceutic (China), MK-8776 from Selleck Chemicals, CHK2 inhibitor II from Abcam. Gemcitabine was dissolved in PBS. Cisplatin, MK-8776 and CHK2 inhibitor II were dissolved in DMSO and used at the specified concentrations.

Dai C.H., Wang Y., Chen P., Jiang Q., Lan T., Li M.Y., Su J.Y., Wu Y, & Li J. (2017). Suppression of the FA pathway combined with CHK1 inhibitor hypersensitize lung cancer cells to gemcitabine. Scientific Reports, 7, 15031.

+ Open protocol

+ Expand

Molecular Mechanism Regulation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

N-Hydroxyphthalimide was purchased from Accela ChemBio Co., Ltd. (Shanghai, China). Propidium iodide (PI), RNase A and 4,6-diamidino-2-phenylindole (DAPI) were from Sigma-Aldrich. Antibodies of mTOR, Phospho-mTOR (Ser2448), Phospho-mTOR (Ser2481), S6K1, Phospho-S6K1 (Thr389), Phospho-S6 Ribosomal Protein (Ser235/236), 4E-BP1, Phospho-4E-BP1 (Ser65), Phospho-Akt (Ser473), Phospho-Akt (Thr308), Cleaved PARP, Cleaved Caspase 3, Caspase 9, cyclin B1, cdc2 were obtained from Cell Signaling Technology; antibodies of S6, Akt, P-ERK1/2, β-actin, Caspase 3, Caspase 8, Bcl-xL, survivin were from Santa Cruz; antibody of ERK was from Epitomics; antibody of eIF4E was obtained from BD Biosciences; Alexa Fluor® 647 donkey anti-mouse IgG antibody was purchased from Invitrogen; all the other secondary antibodies were from Sigma-Aldrich.

Wang M., Zhou A., An T., Kong L., Yu C., Liu J., Xia C., Zhou H, & Li Y. (2016). N-Hydroxyphthalimide exhibits antitumor activity by suppressing mTOR signaling pathway in BT-20 and LoVo cells. Journal of Experimental & Clinical Cancer Research : CR, 35, 41.

+ Open protocol

+ Expand

Protein Expression Analysis in PANC-1 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Denaturing SDS-PAGE sample buffer was utilized for PANC-1 cell lysis through standard methods. Then, separation of protein lysates was done with 10% SDS-PAGE, and they were transferred onto nitrocellulose membranes. TBS containing 0.1% Triton X-100 and 5% nonfat milk was used overnight at 4°C to block the membranes, followed by incubation at 4°C overnight with primary antibodies of STAT3, p-STAT3, JAK, p-JAK, Bcl-2, Bax, caspase-3, caspase-9, and β-actin (Santa Cruz Biotech, Santa Cruz, CA, USA). Upon washing, the membranes were subjected to incubation with HRP-conjugated secondary antibodies for 2 h at room temperature. Detection of the signal was accomplished with the ECL reagents.

Xing H.B., Tong M.T., Wang J., Hu H., Zhai C.Y., Huang C.X, & Li D. (2018). Suppression of IL-6 Gene by shRNA Augments Gemcitabine Chemosensitization in Pancreatic Adenocarcinoma Cells. BioMed Research International, 2018, 3195025.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!