Tet (purity 99.1%) was purchased from Alphabio Biotechnology Co. Ltd (Tianjin, China). The HT-29 cell line was obtained from Cell Bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China). DMEM was purchased from Corning Cellgro Inc. (Herndon, VA, U.S.A.) and the fetal bovine serum (FBS) was obtained from Biological Industries Technologies (Kibbutz Beit Haemek, Israel). DMSO and MTT were acquired from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Trypsin-EDTA solution, penicillin-streptomycin solution, mitochondrial membrane potential (MMP) assay kit with JC-1, caspase 3, 8 activity assay kit, and propidium iodide (PI)/RNase staining solution were obtained from Beyotime Biotechnology Co., Ltd. (Shanghai, China). FITC goat anti-rabbit IgG and Annexin V-FITC Apoptosis Detection Kit were acquired from Tianjin Sungene Biotech Co. Ltd. (Tianjin, China). Anti-Bax, anti-Bcl-2, anti-caspase 3, anti-caspase 8, anti-PARP, anti-cyclin D1 (anti-CCND1), anti-cyclin-dependent kinase 4 (anti-CDK4), anti-phosphorylated Rb (anti-p-Rb) (Ser780), and β-actin antibodies were purchased from Bioss Biotechnology Co. Ltd. (Beijing, China).
Anti caspase 3
Manufactured by Bioss Antibodies
Sourced in China
Anti-caspase 3 is an antibody product provided by Bioss Antibodies. It is used to detect the presence and levels of caspase-3, a key enzyme involved in the process of apoptosis or programmed cell death.
Automatically generated - may contain errors
Lab products found in correlation
Anti bax, by Bioss Antibodies (2 mentions)
Anti bcl 2, by Bioss Antibodies (2 mentions)
Anti bax, by Boster Bio (2 mentions)
Anti caspase 9, by Bioss Antibodies (2 mentions)
Anti caspase 12, by Bioss Antibodies (2 mentions)
Anti grp94, by Boster Bio (2 mentions)
Anti bcl 2, by Wanlei (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Trypsin edta solution, by Beyotime (1 mentions)
Mitochondrial membrane potential assay kit with jc 1, by Beyotime (1 mentions)
β actin antibody, by Bioss Antibodies (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Beyotime (1 mentions)
Annexin 5 fitc apoptosis detection kit, by Sungene Biotech (1 mentions)
Dimethyl sulfoxide (dmso), by Beyotime (1 mentions)
Penicillin streptomycin solution, by Beyotime (1 mentions)
Ab131368, by Abcam (1 mentions)
Gel imaging system, by Thermo Fisher Scientific (1 mentions)
Anti p53, by Cell Signaling Technology (1 mentions)
Enhanced chemiluminescence kit, by GE Healthcare (1 mentions)
8 protocols using anti caspase 3
1
Tet-induced Apoptosis and Cell Cycle Arrest in HT-29 Cells
2
MAP Kinase Pathway Protein Profiling
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The cells were treated with MAPK or ERK inhibitor for 24 h and 5×105 cells were collected for Western blot analysis. Protein (20 μg) was extracted from the treated cells, run on sodium-dodecyl sulfate-polyacrylamide gels (10%), and transferred to nitrocellulose membranes. The membranes were blocked by 5% non-fat milk for 2 h at room temperature. The following primary antibodies were used for overnight incubation at 4°C: anti-caspase-3 (1: 1000, bs-0081R, Bioss, Beijing, China), anti-P53 (1: 2000, Cell Signaling), anti-p-MAPK14 (1: 1000, bs-5476R, Bioss, Beijing, China), anti-MAPK14 (1: 1000, bs-28027R, Bioss, Beijing, China), anti-MEK1 (1: 1000, ABclonal), and anti-p-MEK1 (1: 1000, ABclonal). After washing, the membranes were incubated with the secondary antibody (1: 100, ab131368, Abcam) for 2 h at room temperature. An Enhanced Chemiluminescence kit (No. RPN2133; GE Healthcare Life Sciences, Chicago, IL, USA) was added to the membrane prior to visualization with a gel imaging system (Thermo Fisher Scientific).
3
Histological Analysis of Mouse Embryonic Skull Development
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Mouse embryos were harvested from timed pregnant females. Skulls of collected embryos were fixed in 4% Paraformaldehyde at 4°C for 24 h, then decalcified with formic acid for 14 days. Heads were sectioned (7 μm thickness) and stained with hematoxylin and eosin (H&E) or Azan. Immunofluorescence was performed in PBS/10% bovine serum albumin using the following antibodies: anti-Runx2 (Abcam,ab76956), anti-Sox9 (Abcam, ab3697; 1:50), anti-Ki67 (BD, 550609; 1:50), anti-Sp7/Osterix (Abcam, ab22552; 1:100), anti-Caspase3 (Bioss, bs-0081R), anti-GFP (Abcam, ab13970), anti-COLX (Abcam, ab58632), anti-COLII (Abcam, ab34712), anti-Caspase3 (Abcam, ab44976), anti-Scleraxis (Santa Cruz, sc-518082), Anti-FGF18 (Santa Cruz, sc-393471), anti-Collagen Type I (Proteintech, 14695-1-AP), anti-IHH (Proteintech, 13388-1-AP), anti-β-catenin (CST, #8480), anti-pErk1/2 (CST, #4370), anti-pP38 (CST, #4511), anti-pJNK (Santa Cruz, sc-6254). For immunofluorescence staining, an appropriate secondary antibody conjugated to a fluorescence probe was added. This was then incubated at room temperature for 1 h, followed by rinsing in PBS. Finally, the samples were mounted in an anti-fading mounting media. Results were obtained using an Olympus BX51 upright microscope (Olympus Optical, Tokyo, Japan).
4
Western Blot Analysis of ER Stress Markers
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Western blot was performed as described previously [14 (link)]. Antibodies for anti-calumenin (1: 500), anti-caspase-3 (1: 500), anti-caspase-9 (1: 500), anti-caspase-12 (1: 500) were obtained from Bioss, anti-CHOP (1: 1000), anti-Bcl-2 (1: 1000) were obtained from Wanleibio, and anti-GRP78 (1: 400), anti-GRP94 (1: 400), anti-Bax (1: 400), anti-ATF-6 (1: 400) were purchased from Boster, and anti-p-PERK (1:750), anti-PERK (1:750), anti-spliced XBP-1 (1:500), anti-unspliced XBP-1 (1:500) were obtained from Abcam. Protein was extracted and mixed in loading buffer, and then equal amounts were fractionated on gel and transferred onto Hybond-C Extra nitrocellulose membrane using a semidry transfer apparatus. Protein was blocked with nonfat dry milk and detected with supersignal west pico chemiluminescent substrate.
5
Protein Expression Analysis in Heart Tissue
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The heart tissues and H9 c2 cells in each group were added with RIPA Lysis Buffer (Beyotime) and homogenised on ice. After centrifugation, the supernatant was taken and denatured by protein-loading buffer. After being electrophoresed, the protein was transferred to the nitrocellulose membrane (IB24001, Thermo Fisher Scientific, Shanghai, China), then blocked with skim milk powder, and incubated with primary antibodies (anti-SOD1 (Bioss), anti-SOD2 (Bioss), anti-Calcipressin 1/DSCR 1 (Bioss), anti-RhoA (Bioss), anti-Calpain 2 (Bioss), anti-Caspase-3 (Bioss), anti-phospho-NFKB p65 (Ser281) (Bioss), and anti-Bcl-2 (Bioss)) overnight. Next day, the corresponding secondary antibody was used for continuous incubation, the images were collected and processed after exposure to enhanced chemiluminescence (ECL), and gapdh/β-actinwas used as an internal control. The protein expression level was analysed with ECL-plus reagent (GE Healthcare, Shanghai, China).
6
Protein Expression Analysis with Western Blot
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Western blot was performed as described previously [13 (link)]. The primary antibodies contain anti-caspase-3 (1:500), anti-caspase-9 (1:500), anti-caspase-12 (1:500) (Bioss), anti-CHOP (1:1000), anti-Bcl-2 (1:1000) (Wanleibio), and anti-GRP78 (1:400), anti-GRP94 (1:400), anti-Bax (1:400) (Boster).
7
Silencing IGFBP3 Expression Assay
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Specific siRNAs for human IGFBP3 (5′-GCACAGAUACCCAGAACUUUU-3′) and scrambled siRNA (5′-UAACGACGCGACGACGUAA-3′) were designed and obtained from Biomics Biotechnologies Co., Ltd. (Nantong Shi, Jiangsu Sheng, China). Lipofectamine 3000 was purchased from Thermo Fisher Scientific (Waltham, MA, USA) for cell transfection. Anti-β-tubulin, anti-Bax, anti-caspase 3, anti-B-cell lymphoma-2 (Bcl-2), and anti-IGFBP3 primary antibodies were purchased from Bioss Biotechnology (Beijing, China).
8
Double Immunofluorescence Staining Protocol
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Immunofluorescence staining was performed as described previously [17] . For double immunofluorescence staining, the sections were incubated for 60 h at 4°C in a solution containing rabbit polyclonal anti-GABA (1 : 3000; Sigma) plus mouse monoclonal anti-CB or mouse monoclonal anti-CR (1 : 1000; SWANT) antibodies; and rabbit polyclonal anti-caspase 3 (1 : 1000; Bioss) plus mouse monoclonal anti-CB (1 : 1000; SWANT) antibodies. After washing in PBS, the sections were treated for 2 h at room temperature with an Alexa Fluor 546-conjugated goat anti-rabbit IgG (H + L) (1 : 1000; Invitrogen) or an Alexa Fluor 488-conjugated goat anti-mouse IgG (H + L) (1 : 1000; Invitrogen) and 4',6-diamidino-2-phenylindole (DAPI), washed with PBS, mounted with Vectashield (Vector Laboratories), and visualised using a Nikon A1 confocal microscope equipped with a 100× objective lens (Nikon).
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