CB2-mid DBT cells were treated with indicated concentration of IR700DX-mbc94, mbc94, without light irradiation for 24 h in a water jacketed incubator. ICG (Sigma-Aldrich) was used as the negative control and Doxorubicin hydrochloride (Fisher Scientific) was used as the positive control. To test the toxicity of the IR700DX-mbc94 in cells that express the CB2 receptor but are not tumoral, CHO-K1/CB2 cells were treated with indicated concentration of IR700DX-mbc94 for 24 h. Cell viability was determined by CellTiter-Glo Luminescent Cell Viability Assay kit (Promega) according to the manufacturer’s instructions.
Doxorubicin hydrochloride
Manufactured by Thermo Fisher Scientific
Sourced in United States
Doxorubicin hydrochloride is a cytotoxic agent used in the treatment of various types of cancer. It is an anthracycline antibiotic that intercalates with DNA and inhibits DNA and RNA synthesis. Doxorubicin hydrochloride is commonly used in the management of solid tumors and hematological malignancies.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Thermo Fisher Scientific (2 mentions)
Celltiter glo luminescent cell viability assay kit, by Promega (1 mentions)
Zoledronic acid, by Merck Group (1 mentions)
Mannose, by Tokyo Chemical Industry (1 mentions)
Rm0029 11h3, by Santa Cruz Biotechnology (1 mentions)
Anti cd206, by Santa Cruz Biotechnology (1 mentions)
Hoechst 33342, by AnaSpec (1 mentions)
Poly d lysine, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Ncrc 10001 50, by Cyagen (1 mentions)
C57bl 6j mice, by Jackson ImmunoResearch (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
C1498, by American Type Culture Collection (1 mentions)
Dspe peg2000, by Avanti Polar Lipids (1 mentions)
1 stearoyl 2 hydroxy sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions)
Caspase glo 3 7 assay, by Promega (1 mentions)
Verapamil hydrochloride, by Merck Group (1 mentions)
Verteporfin, by Merck Group (1 mentions)
14 protocols using doxorubicin hydrochloride
1
Evaluating Cytotoxicity of IR700DX-mbc94
2
Synthesis and Characterization of Lipid-PEG Conjugates
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O-Stearoyl Mannose (M-C18), polyethylene
glycol 2000-hydrazone-C18 (PHC), and polyethylene glycol 2000-amide-C18
(PAC) were synthesized following our previously published methods.31 (link),34 Doxorubicin hydrochloride was from Fisher Scientific Co. (Pittsburgh,
PA). Zoledronic acid, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT), PLGA (752H), and poly-d -lysine were from Sigma-Aldrich
(St. Louis, MO). Mannose was from Tokyo Chemical Industry Co., Ltd.
(Portland, OR). Hematoxylin-eosin (H&E) and anti-CD31 antibody
were from Abcam (Cambridge, MA). Hoechst 33342 was from AnaSpec, Inc.
(Fremont, CA). The 5-bromo-2′-deoxyuridine (BrdU) and primary
BrdU monoclonal antibody were from BD Biosciences (San Jose, CA).
Anti-CD206, RM0029-11H3, and FITC-labeled Anti-CD206 antibody were
from Santa Cruz Biotechnology, Inc. (Dallas, TX). Solvents used in
chemical synthesis were of analytical grade.
glycol 2000-hydrazone-C18 (PHC), and polyethylene glycol 2000-amide-C18
(PAC) were synthesized following our previously published methods.31 (link),34 Doxorubicin hydrochloride was from Fisher Scientific Co. (Pittsburgh,
PA). Zoledronic acid, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT), PLGA (752H), and poly-
(St. Louis, MO). Mannose was from Tokyo Chemical Industry Co., Ltd.
(Portland, OR). Hematoxylin-eosin (H&E) and anti-CD31 antibody
were from Abcam (Cambridge, MA). Hoechst 33342 was from AnaSpec, Inc.
(Fremont, CA). The 5-bromo-2′-deoxyuridine (BrdU) and primary
BrdU monoclonal antibody were from BD Biosciences (San Jose, CA).
Anti-CD206, RM0029-11H3, and FITC-labeled Anti-CD206 antibody were
from Santa Cruz Biotechnology, Inc. (Dallas, TX). Solvents used in
chemical synthesis were of analytical grade.
3
Doxorubicin AML Murine Model
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Doxorubicin hydrochloride was purchased from Fisher Scientific Co. (D4193; purity, >95%). Noncontrolled-rate cell cryopreservation medium was bought from Cyagen Co. (NCRC-10001-50). AML cell line C1498 was purchased from the American Type Culture Collection (ATCC). Luciferase and DsRed tagged C1498 cell line was provided by B. Blazar of the University of Minnesota. The cells were cultured in 90% Dulbecco’s modified Eagle’s medium (Gibco) and 10% fetal bovine serum (Gibco) with penicillin (200 U ml−1) and streptomycin (200 U ml−1) (Gibco). The cells were passaged every 1 to 2 days. C57BL/6J mice (4 to 6 weeks, female) were purchased from the Jackson laboratory. All animal tests complied with the animal protocol approved by the Institutional Animal Care and Use Committee of the University of California, Los Angeles.
4
Liposomal Doxorubicin Formulation and Characterization
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All lipids, dipalmitoylphosphatidylcholine
(DPPC), 1-stearoyl-2-hydroxy-sn-glycero-3-phosphocholine
(MSPC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000), cholesterol,
and hydrogenated-l -α-phosphatidylcholine (HSPC) were
purchased from Avanti Polar Lipid. Doxorubicin hydrochloride was purchased
from Fisher Scientific.
LTSLs and NTSLs were synthesized by
a reverse-phase evaporation method.56 (link)−58 (link) Briefly, 5 mg of lipid
(for LTSLs: DPPC–MSPC–DSPE-PEG2000 = 90:10:4; for NTSLs:
HSPC–CHOL–DSPE-PEG2000 = 75:50:3 in molar ratio)59 (link),60 (link),64 (link),65 (link) was dissolved in 1 mL of chloroform, then dried using nitrogen air
and subsequent vacuum. To form the liposomes, the lipid film was hydrated
with a 300 mM citrate buffer (pH 4.0) for 60 min (at 55 °C for
LTSLs and 60 °C for NTSLs). The liposomes were then extruded
10 times through a 400 nm and a 100 nm polycarbonate membrane to obtain
the desired size. The outside pH of the liposome solution was titrated
to pH 7.5 using 0.5 M sodium carbonate. As a result, a pH gradient
was generated across the lipid bilayer.61 (link)−63 (link) DOX was then added to
the liposome solution at a 1:20 DOX/lipid weight ratio and mixed for
60 min (at 37 °C for LTSLs and 60 °C for NTSLs). The final
product was passed through a PD10 column to remove excess DOX.
(DPPC), 1-stearoyl-2-hydroxy-sn-glycero-3-phosphocholine
(MSPC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000), cholesterol,
and hydrogenated-
purchased from Avanti Polar Lipid. Doxorubicin hydrochloride was purchased
from Fisher Scientific.
LTSLs and NTSLs were synthesized by
a reverse-phase evaporation method.56 (link)−58 (link) Briefly, 5 mg of lipid
(for LTSLs: DPPC–MSPC–DSPE-PEG2000 = 90:10:4; for NTSLs:
HSPC–CHOL–DSPE-PEG2000 = 75:50:3 in molar ratio)59 (link),60 (link),64 (link),65 (link) was dissolved in 1 mL of chloroform, then dried using nitrogen air
and subsequent vacuum. To form the liposomes, the lipid film was hydrated
with a 300 mM citrate buffer (pH 4.0) for 60 min (at 55 °C for
LTSLs and 60 °C for NTSLs). The liposomes were then extruded
10 times through a 400 nm and a 100 nm polycarbonate membrane to obtain
the desired size. The outside pH of the liposome solution was titrated
to pH 7.5 using 0.5 M sodium carbonate. As a result, a pH gradient
was generated across the lipid bilayer.61 (link)−63 (link) DOX was then added to
the liposome solution at a 1:20 DOX/lipid weight ratio and mixed for
60 min (at 37 °C for LTSLs and 60 °C for NTSLs). The final
product was passed through a PD10 column to remove excess DOX.
5
Doxorubicin-Induced Apoptosis Assay
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Cells were plated in quadruplicate on two separate 96-well plates at a density of 5000–10,000 cells/well and allowed to incubate overnight (~ 16 h). Additional cells were plated in two sets of triplicates for DOX(+) and DOX(−) control to verify apoptosis induction. Then, 100 μM doxorubicin (DOX) stock solution (prepared from doxorubicin hydrochloride; Fisher Scientific, Cat #ICN15910101) was diluted to 1 μM in media and added to each test well and DOX(+) control wells; 100 μL fresh media was added to DOX (−) control wells. At 10 h and 24 h post DOX treatment, 100 μL of caspase 3/7 GLO reagent (Promega caspase 3/7 GLO Assay) was added to each test well, incubated at room temperature for 15 min, and read on the Wallac Victor2 1420 Multilabel Counter using the luminometer setting.
6
Verteporfin and Verapamil Cytotoxicity Assay
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Verteporfin and (±)-verapamil hydrochloride were purchased from Sigma (St. Louis, MO, USA). Hoechst 33342 and propidium iodide were purchased from Invitrogen Corporation (Waltham, MA, USA). Doxorubicin hydrochloride was purchased from Fisher Scientific (Pittsburgh, KS, USA). The SMARTPool siRNAs targeting YAP1 and control siRNA were purchased from Thermo Scientific Dharmacon (Pittsburgh, PA, USA). The YAP and YAP S127A plasmid DNA were purchased from Addgene (Cambridge, MA, USA).
7
Cyclodextrin-Based Drug Delivery System
8
Chemical Compounds for Cell Biology
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N-(p-amylcinnamoyl)anthranilic acid (ACA), maintained as a 50 mM stock solution in dimethylsulfoxide (DMSO), 2-aminoethoxydiphenyl borate (2-APB; 75 mM stock solution in DMSO), aristolochic acid (75 mM stock solution in DMSO) and 3-methyladenine (3-MA; 50 mM stock solution in DMSO) were purchased from Sigma (St. Louis, MO, USA). N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG; 0.5 M stock solution in DMSO) was purchased from AccuStandard (New Haven, CT, USA). Doxorubicin hydrochloride and tamoxifen citrate were purchased from Thermo Scientific (Waltham, MA, USA). Q-VD-OPh was purchased from R&D Systems (Minneapolis, MN, USA) and maintained as a 50 mM stock solution in DMSO. Propidium iodide (PI) (10 mg/ml solution) was purchased from Thermo Scientific. ApoScreen Annexin V-fluorescein isothiocyanate (FITC) was purchased from Southern Biotech (Birmingham, AL, USA).
9
Radiolabeled Antibody Conjugate Synthesis
10
Hyaluronic Acid-Based Nanoparticle Formulation
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High molecular
weight hyaluronic acid was purchased from Glycosan
Biosystems (Salt Lake, UT). Bifunctionalized poly(ethylene glycol)
with thiol and amine functionalizations, N-(3-(dimethylamino)propyl)-N′-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), sodium borohydride, deuterium
oxide (D2O), and bovine hyaluronidase were purchased from
Sigma-Aldrich; doxorubicin hydrochloride, auric chloride (HAuCl3), and adipic dihydrazide were purchased from Fischer Scientific.
Dulbecco’s modified Eagle’s medium (DMEM) with 4.5 and
1 mg/mLl -glutamine, RPMI 1640 medium with l -glutamine,
fetal bovine serum (FBS) were purchased from Worldwide Medical Supplies.
CD44-directed DNA aptamer and the randomized version of the aptamer
sequence were purchased from Integrated DNA Technologies (IDT). The
Milli-Q water used in all experiments was obtained from a three-stage
Millipore Milli-Q plus 185 purification system (Millipore Corporation),
with a resistivity greater than 18.2 MΩ cm.
weight hyaluronic acid was purchased from Glycosan
Biosystems (Salt Lake, UT). Bifunctionalized poly(ethylene glycol)
with thiol and amine functionalizations, N-(3-(dimethylamino)propyl)-N′-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), sodium borohydride, deuterium
oxide (D2O), and bovine hyaluronidase were purchased from
Sigma-Aldrich; doxorubicin hydrochloride, auric chloride (HAuCl3), and adipic dihydrazide were purchased from Fischer Scientific.
Dulbecco’s modified Eagle’s medium (DMEM) with 4.5 and
1 mg/mL
fetal bovine serum (FBS) were purchased from Worldwide Medical Supplies.
CD44-directed DNA aptamer and the randomized version of the aptamer
sequence were purchased from Integrated DNA Technologies (IDT). The
Milli-Q water used in all experiments was obtained from a three-stage
Millipore Milli-Q plus 185 purification system (Millipore Corporation),
with a resistivity greater than 18.2 MΩ cm.
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