PLGA 50:50 (molecular weight 40–75 kDa), polyvinyl alcohol (molecular weight 30 kDa), 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT), TF, EGCG, CDDP, propidium iodide, and β-actin (clone AC-74) were purchased from Sigma-Aldrich (St Louis, MO, USA). Caspase-3, caspase-9, cytochrome C, p-NF-κB, p-IκBα, p53, and Bcl-2 antibodies were sourced from Cell Signaling Technology (Beverly, MA, USA) while Bax antibody was obtained from Santa Cruz Biotechnology Inc (Santa Cruz, CA, USA). The rabbit anti-mouse and goat anti-rabbit horseradish peroxidase-conjugated secondary antibodies were obtained from Bangalore Genei (Bangalore, India). The polyvinylidene fluoride membrane was obtained from Millipore (Bedford, MA, USA). Caspase-3 inhibitor z-DEVD-fmk was purchased from Calbiochem (Boston, MA, USA). Fetal bovine serum, Dulbecco’s Modified Eagle’s Medium, and Roswell Park Memorial Institute medium were supplied by Invitrogen (Invitrogen, Carlsbad, CA, USA) and the antibiotics by Gibco (Lifetech, Karlsruche, Germany). 2′,7′-dichlorofluorescein diacetate (DCF-DA) and rhodamine 123 from BD Pharmingen (San Diego, CA, USA) were used. Other chemicals used were of analytical grade and sourced locally.
Bcl 2 antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Bcl-2 antibody is a laboratory reagent used to detect the presence and expression levels of the Bcl-2 protein. Bcl-2 is a key regulator of apoptosis, or programmed cell death, and plays a crucial role in cellular survival and proliferation. This antibody can be used in various experimental techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the expression and localization of the Bcl-2 protein in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Bax antibody, by Cell Signaling Technology (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Propidium iodide, by Merck Group (2 mentions)
Cytochrome c, by Cell Signaling Technology (2 mentions)
Cleaved caspase 3 antibody, by Cell Signaling Technology (2 mentions)
Rhodamine 123, by BD (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Bax antibody, by Santa Cruz Biotechnology (1 mentions)
Polyvinyl alcohol (pva), by Merck Group (1 mentions)
Epigallocatechin gallate (egcg), by Merck Group (1 mentions)
P iκbα, by Cell Signaling Technology (1 mentions)
Plga 50 50, by Merck Group (1 mentions)
β actin clone ac 74, by Merck Group (1 mentions)
P nf κb, by Cell Signaling Technology (1 mentions)
Hank s balanced salt solution, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Cleaved parp, by Cell Signaling Technology (1 mentions)
Mtt reagent, by Merck Group (1 mentions)
Immobilon reagent, by Merck Group (1 mentions)
12 protocols using bcl 2 antibody
1
PLGA-based Nanoformulations for Cancer Therapy
2
Mitochondrial Dysfunction Assay Protocol
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MTT reagents and propidium iodide (PI) were obtained from Sigma-Aldrich (St. Louis, MO). The CellROX Green reagent, DMEM, FBS, trypsin, penicillin–streptomycin and Hanks Balanced Salt Solution were purchased from Thermo Fisher Scientific (Waltham, MA); Lipofectamine was obtained from Invitrogen (Carlsbad, CA). The Lowery assay reagent was from BioRad (Hercules, CA). The immunoblot PVDF (polyvinylidene difluoride) membrane and Immobilon reagent were both purchased from Millipore (St. Louis, MO). The Mitochondrial Membrane Potential kit and the ATP Assay kit were purchased from Cayman (Ann Arbor, MI). The Lactate Assay and Mitochondrial Isolation kits along with the PI/RNase solution were purchased from Abcam (Cambridge, MA). Anti-AMPK, p-AMPK (Thr172), FOXO3a, p-FOXO3a (Ser413) and MnSOD p-ATM, γ-H2AX, p-p53, Bim, Bid, Bax, cleaved-PARP, cleaved-caspases 3, 8, and 9, cytochrome C and Bcl-2 antibodies and the secondary anti-mouse IgG-horseradish peroxidase antibody were obtained from Cell Signaling (Danvers, MA).
3
Antibody Panel for Protein Analysis
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The antibodies used in this study include Nampt antibody (abcam, Cambridge, MA), FoxO1 antibody (Santa Cruz Biotechnology Inc, Santa Cruz, CA), GAPDH antibody (Sigma-Aldrich, St. Louis, MO), thioredoxin1 antibody (generated by this laboratory), Ac-FoxO1 antibody, Akt antibody, phospho-specific Akt antibody, ERK1/2 antibody, phospho-specific ERK1/2 antibody, STAT3 antibody, phospho-specific STAT3 antibody, Bcl2 antibody, Bcl-xL antibody, Bax antibody (Cell Signaling Technology, Danvers, MA), and MnSOD antibody (BD Transduction Laboratory, San Jose, CA). Ly.6B.2 antibody (AbD Serotec, Raleigh, NC)
4
Apigenin-Induced Senescence Cell Analysis
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Apigenin, Senescence Cells Histochemical Staining Kit, Griess reagent were purchased from Sigma Chemicals Co., USA. Dulbecco's modified Eagle's medium (DMEM) and Roswell Park Memorial Institute 1640 medium (RPMI-1640) supplemented with l -glutamine, fetal bovine serum (FBS), penicillin, streptomycin, Dulbecco's phosphate-buffered saline (D -PBS) and Hank's balanced salt solution (HBSS) were all procured from Gibco (Invitrogen), USA. JC-1 fluorescent dye was obtained from Life Technologies (Thermo Fisher Scientific Corporation, Carlsbad, CA, USA). Phospho-Rb (Ser780) Antibody, Bax Antibody, Bcl-2 Antibody, Anti-mouse IgG and Anti-rabbit IgG were procured from Cell Signaling Technology®, USA while Anti-p21WAF1 (Ab-1) was obtained from Calbiochem®, Darmstadt, Germany. Cyclin D1, Cyclin E, p53, p16 antibodies were procured from Santa Cruz Biotechnology, Inc., Dallas, USA while β-actin antibody was obtained from Sigma Chemicals Co., USA. JC-1 fluorescent dye was obtained from Life Technologies (Thermo Fisher Scientific Corporation, Carlsbad, CA, USA). All other chemicals used were of the highest analytical grade available. The chemicals were used as obtained without further purification. Milli-Q water obtained from Milli-Q Integral 3 system (Merck Millipore, Germany) was used for all experiments.
5
Immunoblotting Workflow for Angiogenesis Proteins
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Bcl-xL antibody (1:1000, Cat#: ab32370) and VASH1 (1:1000, Cat#: ab199732) were purchased from Abcam. Bcl-2 antibody (1:1000, Cat #: 15071S), and Cleaved Caspase-3 (1:1000, Cat#: 9661S) were purchased from Cell Signaling Technologies. α-tubulin (1:1000, Cat#: T6199), FLAG (1:1000, Cat#: F3165), Tyr-Tub antibody (1:1000, Cat# MAB1864-I), and VASH2 (1:1000, Cat#: MABC536) were purchased from Sigma. GAPDH (1:5000, Cat#: sc-32233) was purchased from Santa Cruz. SVBP (CCDC23, 1:1000, Cat#: PA5-52569) was purchased from Invitrogen. The deTyr-Tub antibody was developed by Takashi Hotta and Ryoma Ohi [17 (link)]. This antibody is now commercially available through RevMab under catalogue number RM444 (1:10000). Alexa Fluor 568 (1:1000, Cat#: A11011), Alexa Fluor 594 (1:1000, Cat#: A11012) and Hoechst 33258 (1:5000, Cat#: H3569) were purchased from Invitrogen (Waltham, MA, USA).
6
Western Blot Analysis of Bcl-2 and Bax in Cardiomyocytes
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Proteins were extracted from cardiomyocytes or heart tissues with RIPA lysis buffer (Beyotime Biotechnology). Protein samples (60 μg) were separated by SDS‐PAGE electrophoresis and transferred to PVDF membranes (Millipore). After blocking, the protein on the membrane was incubated with primary antibody Bcl‐2 antibody (Cell Signaling Technology) and Bax antibody (Cell Signaling Technology) at 4°C overnight, and then incubated with HRP‐conjugated secondary antibody the next day. Then FluorChemE imager (Alpha) was used for visualization, and the expression level of specific protein was normalized to GAPDH level.
7
Breast Cancer Drug Resistance Study
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MCF-7/ADM and MCF-7 cell lines were purchased from Aolushengwu (https://aolushengwu.biomart.cn/ , Shanghai, China). Human breast cancer azithromycin-resistant cell lines (MCF-7/ADM) were purchased from Shanghai Zhen Biotechnology (Shanghai, China). miR-93 mimics, inhibitor, and primers were purchased from Thermo Fisher Scientific, Inc. Waltham, MA, USA). Adriamycin, fetal bovine serum, thiazolyl (MTT) and RPMI-1640 medium were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dimethyl sulfoxide (DMSO), reverse transcription kit, real-time-quantitative PCR (RT-qPCR) kit, Bcl-2 antibody, P-gp antibody, GAPDH antibody, β-actin were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). TRIzol kit, liposomes (Lipofectamine 2000), U6 snRNA real-time PCR kit which were purchased from Invitrogen (Thermo Fisher Scientific, Inc., Carlsbad, CA, USA).
The study was approved by the Ethics Committee of Shandong Provincial Hospital affiliated to Shandong University (Jinan, China).
The study was approved by the Ethics Committee of Shandong Provincial Hospital affiliated to Shandong University (Jinan, China).
8
Apoptosis Induction by Paris Saponins
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Paris saponin I, II, VI, and VII were purchased from the ZheJiang Institute for Food and Drug Control (batch no. 111590,111591,111592,111593, Hangzhou, China). Gefitinib was obtained from Tocris Bioscience (Cat. No.3000, Avonmouth, Bristol, United Kingdom) The purity was greater than 99% and dissolved in dimethyl sulfoxide (DMSO) then stored at −20°C. The drugs were diluted in Dulbecco’s modified Eagle’s minimum essential medium (DMEM) to achieve the final concentration used for the following experiment. We used DMEM and fetal calf serum (Hyclone Co., Logan, UT, USA); FITC Annexin-V Apoptosis Detection kit (BD Biosciences, NJ, USA); as well as Hoechst (Sigma-Aldrich, St. Louis, MO, USA); rabbit anti-rat PI3 Kinase p110α, AKT antibody, Phospho-Akt Antibody, Bcl-2 Antibody, Bax Antibody, Caspase-3 at 1: 1000 dilution (Cell Signaling Technology, Danvers, MA, USA); and a monoclonal mouse anti-rat caspase-9 at 1:1000 dilution (Cell Signaling Technology, Danvers, MA, USA).
9
Western Blot Antibody Validation
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Experiments were conducted using the following primary antibodies: Flag-tag antibody (Cell Signaling Technology, 8146 T, 1:1000 dilution), HA-tag antibody (Proteintech, 66006, 1:5000 dilution), caspase 3 antibody (Cell Signaling Technology, 9662 S, 1:1000 dilution), cleaved caspase 3 antibody (Cell Signaling Technology, 9661 S, 1:1000 dilution), cleaved PARP antibody (Cell Signaling Technology, 5625 S, 1:1000 dilution), β-actin antibody (Proteintech, 66009-1-Ig, 1:5000 dilution), Bax antibody (Cell Signaling Technology, 2772 S, 1:1000 dilution), BCL-2 antibody (Cell Signaling Technology, 15071, 1:1000 dilution), p53 antibody (Abcam, ab26, 1:1000 dilution) for p53-DBD, GST tag antibody (Cell Signaling Technology, 2625 S, 1:1000 dilution), and His-tag antibody (Abbkine, ABT2050, 1:5000 dilution). The secondary antibodies HRP-conjugated goat anti-mouse IgG (Abbkine, A21010, ATSDE1601, 1:2000 working dilution) and HRP-conjugated goat anti-rabbit IgG (Absin, abs20040, AS004, 1:2000 working dilution) were used. These antibodies were validated by western blotting according to the manufacturer’s website. Bands were visualized by enhanced chemiluminescence detection reagents (Vazyme, E411-04). Full blots have been included in the Source data file.
10
Protein Extraction and Western Blot Analysis
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Cells were lysed either in Flag-lysis buffer or directly solubilized in 1× SDS buffer, then subjected to SDS-PAGE. Cells lysed with Flag-lysis buffer were set on ice for 30 min, and then centrifuged at 20,000 × g for 10 min. The supernatants were then mixed with 4× SDS buffer and subjected to SDS-PAGE. The antibodies used in this research were: PDE3A antibody from Bethyl Laboratories (1:1000, Cat# A302-740A); SLFN12 antibody from abcam (1:400, Cat# ab234418); Cleaved-Caspase-3 antibody from Cell Signaling technology (1:1000, Cat# 9661); Caspase-9 antibody from Cell Signaling technology (1:1000, Cat# 9502); PARP antibody from Cell Signaling technology (1:1000, Cat# 9542); Bcl-2 antibody from Cell Signaling technology (1:1000, Cat# 4223); anti-Rabbit-HRP antibody from Sigma-Aldrich (1:5000, Cat# A0545); anti Mouse-HRP antibody from Sigma-Aldrich (1:5000, Cat# A9044); MYC-HRP antibody from MBL (1:1000, M-047-7); Flag-HRP antibody from Sigma-Aldrich (1:10,000, Cat# A8592); Actin-HRP antibody from MBL (1:50,000, Cat# PM053-7); α-Tubulin-HRP antibody from MBL (1:50,000, PM054-7); and anti-GAPDH-HRP antibody from MBL (1:50,000, Cat# M171-1).
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