Simvastatin was purchased from Shanghai Yanjing Biological Technology Co., Ltd. (Shanghai, China), nitro tetrazolium chloride (NBT) was purchased from Shanghai Haoran Bio Technologies Co., Ltd. (Shanghai, China), caspase-3 rabbit polyclonal antibody was purchased from Cell Signaling Technology, Inc. (Danvers, USA; cat. no. 9662), and SABC immunohistochemical staining kits were purchased from Shanghai S&S Bio & Tech Co., Ltd. (Shanghai, China; http://www.ssmotor-sh.com/ ), upgraded packing of one step TUNEL apoptosis in situ assay kits were purchased from Jiangsu Keygen Biotech Co., Ltd. (Jiangsu, China). ECL luminescent liquids was purchased from Cell Signaling Technology Inc.).
Caspase 3 rabbit polyclonal antibody
Manufactured by Cell Signaling Technology
Sourced in United States
Caspase-3 rabbit polyclonal antibody is a laboratory reagent used to detect the presence and expression levels of caspase-3 protein. Caspase-3 is a key executioner enzyme involved in the process of apoptosis, or programmed cell death. This antibody can be used to assess caspase-3 activation and its role in various biological processes and disease states.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by Merck Group (1 mentions)
T per lysis buffer, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Picoepd western reagent kit, by Elpis Biotech (1 mentions)
β actin antibody, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Roche (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
4 protocols using caspase 3 rabbit polyclonal antibody
1
Simvastatin-Induced Apoptosis Assay
2
Protein Extraction and Immunoblotting for Apoptosis Markers
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Rat lung tissues were homogenized in cold T-PER™ lysis buffer (Pierce Biotechnology, Inc., Rockford, IL, USA) with a protease inhibitor cocktail (Roche Diagnostics GmbH). Subsequently, the lysates were quickly sonicated three times for 15 sec in an ice bath, boiled for 5 min at 95°C and stored at −80°C prior to use. Aliquots of proteins were separated by 10% sodium dodecyl sulfate polyacrylamide gel, transferred onto a polyvinylidene difluoride membrane and detected using antibodies against LC3B(D11) XP® rabbit monoclonal antibody (1:1,000; #3868s; Cell Signaling Technology, Inc.), Beclin-1 (H-300) rabbit polyclonal antibody (1:1,000; sc-11427; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) or caspase-3 rabbit polyclonal antibody (1:1,000; #9662s; Cell Signaling Technology, Inc.), respectively. β-actin (1:5,000; Sigma Aldrich, Shanghai, China) was used as a loading control.
3
Protein Expression Quantification by Western Blot
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Whole cell lysates were prepared using a lysis buffer (10 mM Tris-Cl, pH 7.4, 120 mM NaCl, 25 mM KCl, 2 mM EGTA, 1 mM EDTA, 0.5% Triton X-100, and protease inhibitor cocktail). Aliquots of lysates were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under protein denaturing conditions. Proteins separated were transferred to polyvinylidene fluoride membranes, which were incubated with an appropriate primary antibody overnight at 4°C and then with a secondary antibody conjugated to horseradish peroxidase for 1 h at room temperature. Immunoreactive bands were detected using a picoEPD Western Reagent kit (ELPIS-Biotech, Daejeon, Korea) and subjected to densitometric analysis. The primary antibody for MMP-1 was purchased from Calbiochem (San Diego, CA, USA). Rabbit polyclonal caspase-3 antibody and rabbit polyclonal caspase-9 were purchased from Cell Signaling (Danvers, MA, USA). The mouse monoclonal β-actin antibody was from Sigma-Aldrich (St. Louis, MO, USA).
4
Protein Expression Analysis of LF Tissue
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The sliced LF tissues and cells were washed 3 times with PBS and dissolved in RIPA lysis buffer with a protease inhibitor and EDTA (Roche, Grenzach, Germany) to prepare protein homogenates. Equal amounts of protein from LF tissue or cells were separated by 12.5% or 10% SDS-PAGE according to the kDA of the targeted protein and transferred to 0.22-μm polyvinylidene fluoride (PVDF) membranes (Merck Millipore, CA, U.S.A). The membranes were blocked with 5% fat free milk at room temperature (RT) for 1 h and then incubated with primary antibodies (all at 1:1000 dilution) overnight at 4°C, including rabbit monoclonal anti-EDG2 (Abcam, Cambridge, UK), rabbit polyclonal Caspase 3 antibody (Cell Signaling Technology, Danvers, MA, U.S.A), rabbit monoclonal anti-phospho-Akt (Ser308) (CST), rabbit monoclonal anti-phospho-Akt (Ser473) (CST), rabbit monoclonal anti-Akt1 (CST), rabbit monoclonal anti-cdc2 (CST), rabbit monoclonal anti-CyclinB1 (CST) and rabbit monoclonal anti-GAPDH (CST). After 3 washes with Tris-buffered saline containing Tween-20 (TBST), the membranes were incubated with anti-rabbit IgG conjugated with IRDye 800CW (CST) for 1 h at RT. After 3 washes in TBST, the immunoreactive bands were detected using the Odyssey infrared imaging system (LI-COR). Positive immunoreactive bands were quantified by Image-Pro Plus 6.0 (IPP6.0) software and normalized to GAPDH.
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