Cell lysates were prepared for immunoblotting, using antibodies against cullin1 (Santa Cruz, Dallas, Texas), phospho-H2AX at Ser139 (γH2AX), ATG7, cleaved caspase 3, cleaved PARP, Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit, Pro-Survival Bcl-2 Family Antibody Sampler Kit, IAP Family Antibody Sampler Kit, ORC1, CDT1 (Cell Signaling, Boston, MA), tubulin (Likun Trade Co., China), NOXA (Millipore, Billerica, MA) and LC3 (Sigma, St. Louis, MO).
Pro apoptosis bcl 2 family antibody sampler kit
Manufactured by Cell Signaling Technology
Sourced in United States
The Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit contains a set of antibodies that target various pro-apoptotic Bcl-2 family proteins. The kit includes primary antibodies specific to Bax, Bak, Bid, Bad, and Bim. These antibodies can be used to detect and study the expression and localization of these pro-apoptotic regulators in cellular and biochemical assays.
Automatically generated - may contain errors
Lab products found in correlation
Iap family antibody sampler kit, by Cell Signaling Technology (3 mentions)
Pro survival bcl 2 family antibody sampler kit, by Cell Signaling Technology (3 mentions)
Cullin1, by Santa Cruz Biotechnology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Glut1 primary antibody, by Abcam (1 mentions)
Anti beta actin primary antibody, by Santa Cruz Biotechnology (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Apoptosis western blot cocktail, by Abcam (1 mentions)
Mouse anti caspase 8 monoclonal antibody, by Cell Signaling Technology (1 mentions)
Carfilzomib, by Selleck Chemicals (1 mentions)
Thenoyltrifluoroacetone, by Merck Group (1 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Oligomycin, by Merck Group (1 mentions)
Advanced dmem f12, by Thermo Fisher Scientific (1 mentions)
B27 supplement, by Thermo Fisher Scientific (1 mentions)
α tubulin, by Merck Group (1 mentions)
Parp antibody, by Cell Signaling Technology (1 mentions)
Anti ubiquitin antibody, by Merck Group (1 mentions)
Gapdh antibody, by Santa Cruz Biotechnology (1 mentions)
Caspase 3 antibody, by Cell Signaling Technology (1 mentions)
7 protocols using pro apoptosis bcl 2 family antibody sampler kit
1
Immunoblotting Analysis of Cellular Signaling
2
Regulation of Cell Cycle and Apoptosis
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Kyse450 and TE1 cells were transfected with siControl or siROC1 for 96 h and then proteins were collected for western blot analysis, using antibodies against p21, p27, WEE1, pH3, cleaved caspase-3, cleaved poly (ADP) ribose polymerase (PARP), Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit, Pro-Survival Bcl-2 Family Antibody Sampler Kit, IAP Family Antibody Sampler Kit and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Cell Signaling, Boston, MA). Densitometric analysis for the quantification relative to GAPDH was performed using the Image J software.
3
Protein Expression Analysis of PCa Cells
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PCa cells seeded in 100-mm cell culture dishes were treated with Etn for different time points. After treatment, cells were subjected to immunoblotting as described previously 33 (link). Lysates from fresh tumor samples were prepared using BeadBlaster microtube homogenizer (LabRepCo) following the manufacturer's guidelines. Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then were transferred onto polyvinylidene difluoride (PVDF) membranes. Membranes were incubated with anti-light chain 3 primary antibody (LC3; Cell Signaling Technologies, #2775S, 1:500) or GLUT1 primary antibody (Abcam, #ab115730, 1:1000) overnight at 4 °C; anti-beta-actin primary antibody (Santa Cruz Biotechnology, #SC-47778, 1:1000) was used as a loading control. Subsequently, membranes were incubated with goat anti-mouse (Santa Cruz Biotechnology, #sc-516102, 1;8000) or goat anti-rabbit (Santa Cruz Biotechnology, #sc-2357, 1:8000) IgG horseradish peroxidase-conjugated secondary antibodies. All antibodies against pro-apoptotic proteins were from Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit (Cell Signaling Technologies, #9942T). The signal was visualized using the Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, #32106). Signal intensities were quantified using ImageJ and were normalized to the respective β-actin signal intensity.
4
Western Blot Analysis of Apoptosis Markers
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Western blot analysis was performed, as described previously (7 (link)) using the following: mouse anti-c-FLIP monoclonal antibody (NF6; cat.no ALX-804-428; Alexis Biochemicals, San Diego, CA, USA), mouse anti-caspase-8 monoclonal antibody (cat.no 9746; Cell Signaling Technology, Inc. Danvers, MA, USA), mouse anti-DR4 monoclonal antibody (cat.no ab13890; Abcam, Cambridge, UK), rabbit anti-DR5 polyclonal antibody (cat.no ab47179; Abcam), Pro-Apoptosis Bcl-2 Family Antibody Sampler kit (cat.no 9942; Cell Signaling Technology, Inc.) and Apoptosis Western Blot Cocktail (cat.no ab136812; Abcam). An enhanced chemiluminescence detection reagent (SuperSignal West Pico) was used for detection (Pierce Biotechnologies, Rockford, IL, USA) and α-tubulin and β-actin were used as loading controls. All western blots were representative of at least three independent experiments.
5
Proteomic Analysis of Mitochondrial Dysfunction
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PARP antibody (Cell Signaling Technology), caspase 3 antibody (Cell Signaling Technology), GAPDH antibody (Santa Cruz Biotechnology), Anti‐Ubiquitin antibody (Sigma‐Aldrich), Anti‐ATP synthase antibody (merckmillipore), OPA1 antibody (Cell Signaling Technology), α‐Tubulin (Sigma‐Aldrich), CTR1 (PROTEINTECH), Pro‐Apoptosis Bcl‐2 Family Antibody Sampler Kit (Cell Signaling Technology) were used. All the inhibitors in this study were used at the indicated concentration unless stated independently: mitochondrial complex III inhibitor antimycin A (sigma A8674; 5 × 10−6m ), mitochondrial complex II inhibitor thenoyltrifluoroacetone (sigma T27006; 2 × 10−6m ), mitochondrial complex V inhibitor oligomycin (sigma 75351; 1 × 10−6m ), mitochondrial complex I inhibitor rotenone (sigma R8875; 5 × 10−9m ), cisplatin (sigma; 20 × 10−6m ), N‐acetyl‐L‐cysteine (sigma A7250; 5 × 10−3m ), MG132 (selleckchem S2619; 150 × 10−9m ), bortezomib (selleckchem S1013; 25 × 10−9m ), carfilzomib (selleckchem PR‐171; 10 × 10−9m ), FCCCP (sigma; 10 × 10−6m ). Corning matrigel matrix bulk (Corning), epidermal growth factor (Thermo), B27 supplement (Gibco), advanced DMEM/F12 (Gibco) were used. The methoxy PEG5000‐ PLGA (MW: 5000, mPEG‐PLGA) polymers were purchased from Xi'an Ruixi Biological Technology Co., Ltd., China.
6
Molecular Apoptosis Profiling in Liver Cells
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HepG2 and Huh7 cell lysates treated with CQ were prepared for western blot analysis, using antibodies against cleaved caspase-3, cleaved poly(ADP-ribose) polymerase (PARP), the Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit, the Pro-Survival Bcl-2 Family Antibody Sampler kit, IAP Family Antibody Sampler kit and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Cell Signaling, Boston, MA, USA).
7
Western Blot Analysis of Cellular Proteins
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Protein samples were separated by one-dimensional 12% Tris–glycine SDS-PAGE, then transferred to nitrocellulose membranes (Bio-Rad, CA, USA). For membrane blocking, membranes were incubated with 1×TBST containing 5% skim milk for 1 h after washing with 1×TBST. nitrocellulose membranes were incubated overnight with the primary antibodies 1:1000 in 5% BSA at 4 ℃ in the dark. After washing three times with TBST for 5 min, 1:5000 horseradish peroxidase-conjugated secondary antibodies (Cell Signaling Technology, MA, USA) were added to 5% skim milk, and membranes were incubated at room temperature for 1 h. Blots were detected using a chemiluminescence SignalFire™ ECL Reagent (Cell Signaling Technology, MA, USA) by Ez-Capture MG system (ATTO, NY, USA), and the relative intensity of the western blot bands was measured using ImageJ 1.53a (National Institutes of Health). GAPDH antibody, purchased from GeneTex (CA, USA), was used as a loading control. The antibodies used in this study included anti-CYP11A1, anti-Snail, anti-Vimentin, anti-Cdk2, anti-Cdk4, anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204), anti-pp44/42 MAPK (Erk1/2), anti-cdc25c, and anti-phospho-cdc25c Ser216. Cell Cycle/Checkpoint Antibody Sampler Kit, Apoptosis Antibody Sampler Kit, and Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit were also purchased from Cell Signaling Technology (MA, USA).
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