Cells were lysed in Ripa buffer containing 150 mM sodium chloride, 1.0% Nonidet P-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, 50 mM Tris (pH 8.0), and a protease inhibitor cocktail (Roche Applied Science, Vienna, Austria). Electrophoresis was performed as previously described [57 (link)]. The following primary antibodies were used for Western blot analysis: anti-poly (ADP-ribose) polymerase (PARP), anti-caspase 3, anti-B-cell lymphoma 2 (Bcl-2), anti-EGR1, anti-β-actin (1:1000; Cell Signaling Technology, Danvers, MA, USA), and anti-GADD45α (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA). Following incubation with the corresponding horseradish peroxidase-conjugated secondary antibodies (1:1000; Santa Cruz Biotechnology), immunoreactive bands were visualized using enhanced chemiluminescence (Bio-Rad Laboratories, Hercules, CA, USA). The original Western Blot pictures can be found in the Supplementary Materials .
Anti b cell lymphoma 2 bcl 2
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-B-cell lymphoma 2 (Bcl-2) is a lab equipment product that detects the Bcl-2 protein, a key regulator of apoptosis (programmed cell death). It is a reliable tool for researchers studying cell survival and death pathways.
Automatically generated - may contain errors
Lab products found in correlation
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Enhanced chemiluminescence, by Bio-Rad (1 mentions)
Anti egr1, by Cell Signaling Technology (1 mentions)
Anti gadd45α, by Santa Cruz Biotechnology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti caspase 3, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Nitrocellulose membrane, by GE Healthcare (1 mentions)
Cytobuster protein extraction reagent, by Merck Group (1 mentions)
Ecl plus western blotting detection reagent, by GE Healthcare (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Anti bax ab32503, by Abcam (1 mentions)
Rpmi 1640 medium, by Vitrocell (1 mentions)
Bicinchoninic acid protein quantification kit, by Beyotime (1 mentions)
Rapamycin, by Merck Group (1 mentions)
Anti p62, by Cell Signaling Technology (1 mentions)
Anti protein kinase b akt, by Cell Signaling Technology (1 mentions)
Antithrombin, by Merck Group (1 mentions)
Anti beclin 1, by Cell Signaling Technology (1 mentions)
5 protocols using anti b cell lymphoma 2 bcl 2
1
Western Blot Analysis of Apoptosis Markers
2
Famitinib Modulates Protein Expression
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Total proteins prior and subsequent to famitinib treatment were extracted from BGC-823 and MGC-803 cell pellets using CytoBuster Protein Extraction Reagent (Merck Millipore, Darmstadt, Germany). Proteins were quantified with a Pierce BCA Protein Assay kit (Thermo Fisher Scientific, Inc.), and ~20 µg of protein was separated on 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and proteins were then transferred to a nitrocellulose membrane (GE Healthcare Life Sciences, Chalfont, UK), which was subsequently incubated with anti-cyclin B1 (dilution, 1:1,000; catalog no., AJ1208a; Abgent Inc., San Diego, CA, USA), rabbit polyclonal anti-B-cell lymphoma 2 (BCL2; dilution, 1:1,000; catalog no., 2872; Cell Signaling Technology, Inc., Danvers, MA, USA) and mouse monoclonal anti-β-actin (dilution, 1:3,000; catalog no., A5441; Sigma-Aldrich, St. Louis, MO, USA) antibodies at 4°C overnight. Secondary anti-rabbit and anti-mouse horseradish peroxidase-conjugated IgG antibodies (dilution, 1:3,000; catalog nos., 7074 and 7076, respectively; Cell Signaling Technology, Inc.) were applied and allowed to incubate at room temperature for 1 h. Proteins were visualized with ECL Plus Western Blotting Detection Reagent (GE Healthcare Life Sciences).
3
Investigating Cancer Inflammation and Immunity Crosstalk
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RPMI-1640 medium, PBS, penicillin-streptomycin antibiotic solution, and trypsin were purchased from Vitrocell Embriolife (Campinas, Brazil). Anti-β-actin (sc-47778), anti-COX-2 (sc376861), and anti-Toll-like receptor 4 (TLR-4) (sc16240) were purchased from Santa Cruz Biotechnology (Irvine, CA, USA); anti-NF-κB (ab13594), anti-IκBα (ab32518), anti-IκBβ (ab7547), anti-TNF-α (ab8348), and anti-BAX (ab32503) were acquired from Abcam (Cambridge, UK) and anti-B-cell lymphoma 2 (BCL-2) was purchased from Cell Signaling (Danvers, MA, US). RNeasy® Micro kit (74004), RT2 First Strand kit (330401) and RT2 Profiler PCR array (PAHS 181ZA-Human Cancer Inflammation and Immunity Crosstalk) were obtained from Qiagen (Hilden, Germany).
4
Autophagy Regulation in Cardiac Injury
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Antithrombin, rapamycin, A6730, Evans blue, and triphenyltetrazolium chloride (TTC) were purchased from Sigma-Aldrich (St. Louis, MO). Anti-beclin-1 (1:1000), anti-GAPDH (1:5000), anti-protein kinase B (Akt; 1:1000), anti-phosphorylated (p)-Akt (Ser473,1:1000), anti-autophagy protein microtubule-associated protein 1 light chain-3B (LC3B; 1:1000), anti-B-cell lymphoma 2 (Bcl-2; 1:1000), anti-Bcl-2 associated x (Bax; 1:1000), and anti-p62 (1:1000) antibodies were obtained from either Cell Signaling Technology (Danvers, MA) or Abcam (Cambridge, UK). Phenylmethane sulphonyl fluoride (PMSF), the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay kit, and the bicinchoninic acid protein quantification kit were obtained from Beyotime (Shanghai, China). The creatine kinase (CK) assay kit was purchased from the Jiancheng Bioengineering Institute (Nanjing, China). Goat anti-rabbit secondary antibodies were purchased from Biosharp (Guangzhou, China).
5
CTRP3 Regulation of Oxidative Stress
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Melatonin and palmitic acid (PA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Recombinant human CTRP3 (rhCTRP3) was purchased from Aviscera Bioscience (Santa Clara, CA, USA). Anti-B-cell lymphoma-2 (Bcl-2, 1:1000) was obtained from Cell Signaling Technology (Danvers, Massachusetts). Anti-Nrf2 (1:1000), anti-heme oxygenase-1 (HO-1, 1:1000), anti-superoxide dismutase 2 (SOD2, 1:1000), anti-tumour necrosis factor-α (TNF-α, 1:200 for staining), anti-CD68 (1:200 for staining) and anti-CTRP3 (1:500 for western blotting, 1:200 for staining) were purchased from Abcam (Cambridge, UK). Anti-adiponectin (1:200 for staining) was purchased from Proteintech (Chicago, IL, USA). The secondary antibody used in the study was purchased from LI-COR Biosciences (1:10,000 dilution). Dihydroethidium (DHE) was purchased from Invitrogen (Carlsbad, CA, USA). 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA), total SOD assay kit, catalase (CAT) assay kit, and glutathione (GSH) assay kit were purchased from the Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The mini-osmotic pump was purchased from Alzet (DURECT Corp, Cupertino, CA). Cell counting kit-8 (CCK-8) was obtained from Dōjindo Laboratories (Kumamoto, Japan). All other chemicals were of analytical grade.
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