The anti-CREB rabbit antibodies (cat# 9197) were obtained from Cell Signaling (Danvers, MA). The BDNF antibody was from Sigma (cat# AV41970, St. Louis, MO) and βIII-Tubulin was from Promega (cat# G7121, Madison, WI). The CRE concensus (cat# sc-2504) and mutant (cat# sc-2517) oligonucleotides were from Santa Cruz Biotechnologies (Santa Cruz, CA). Human recombinant IL-6 and BDNF were from R&D Systems. Colchicine and nocodazole were from Tocris Bioscience; Ciliobrevin D was from EMD Millipore and PGE2 was from Cayman Chemical Company. Capsaicin and lidocaine were from Sigma Aldrich. Stock solutions for Colchicine, nocodazole and Ciliobrevin D were made in cell culture grade 100% DMSO. BDNF and IL-6 stock solution was made in sterile PBS containing 0.1% BSA and TrkB/Fc stock solution was made in sterile PBS. Capsaicin and PGE2 stock solutions were made in 100% ethanol. Lidocaine stock solutions were made in sterile saline. All drugs except were diluted to final concentrations in sterile PBS for injection.
Colchicine
Manufactured by Bio-Techne
Sourced in Japan, United States, United Kingdom
Colchicine is a chemical compound commonly used in research laboratories. It functions as a microtubule-disrupting agent, inhibiting the polymerization of microtubules. This property makes it a valuable tool for studying cellular processes and structures that rely on microtubule dynamics.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Nocodazole, by Bio-Techne (2 mentions)
Vinblastine, by Selleck Chemicals (2 mentions)
Paclitaxel, by Selleck Chemicals (2 mentions)
Alisertib, by Selleck Chemicals (2 mentions)
Rigosertib, by Selleck Chemicals (2 mentions)
Blebbistatin, by Selleck Chemicals (2 mentions)
Abt 751, by Apexbio (2 mentions)
Bi2536, by Selleck Chemicals (2 mentions)
Cisplatin, by Bio-Techne (2 mentions)
Mitoxantrone, by Bio-Techne (2 mentions)
Docetaxel, by Bio-Techne (2 mentions)
Paclitaxel, by Bio-Techne (2 mentions)
Vincristine, by Bio-Techne (2 mentions)
Vinblastine, by Bio-Techne (2 mentions)
Verapamil, by Merck Group (2 mentions)
Fura 2 am, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
15 protocols using colchicine
1
Antibody and Reagent Sourcing for Cell Signaling
2
Small Molecule Inhibitors Protocol
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Rigosertib, vinblastine, blebbistatin, BI2536, alisertib, and paclitaxel were purchased from Selleck chemicals. ABT-751 was purchased from ApexBio. STLC and colchicine were purchased from Tocris. ARS-853 was generously provided by the Shokat lab (UCSF).
3
Karyotyping to Assess Genomic Stability of MSCs
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The genomic stability of AML-MSCs and HD-MSCs was monitored by the G-banded chromosome karyotype analysis. In brief, the aforementioned MSCs in metaphase were treated with colchicine (Tocris) and photographed with an Olympus DA71 microscope (Tokyo) as we recently reported [9 (link), 12 (link), 23 (link)].
4
Small Molecule Inhibitors Protocol
5
Paclitaxel Resistance Mechanism Evaluation
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[3H]-Paclitaxel (37.9 Ci/mmol) was purchased from Moravek Biochemicals, Inc. (Brea, CA). Dulbecco modified Eagle medium (DMEM), fetal bovine serum (FBS), phosphate buffer saline (PBS), 10,000 IU/ml penicillin and 10,000 μg/ml streptomycin, and trypsin 0.25% were purchased from Hyclone (Waltham, MA). Monoclonal antibody against GAPDH was purchased from Cell Signaling Technologies (Beverly, MA). The polyclonal antibody PA5-23652 for ABCC10 was obtained from Thermo scientific (Rockford, IL). Monoclonal antibody 3D7G8 against EphB4 was obtained from Life technologies, Invitrogen (New York, NY). NVP-BHG712 was a gift from Novartis (Basel, Switzerland). Cepharanthine was generously donated by Kakenshoyaku Co. (Tokyo, Japan). PAK-104P was a gift from Nissan Chemical Industries (Tokyo, Japan). Paclitaxel, docetaxel, colchicine, mitoxantrone, vincristine, vinblastine and cisplatin were purchased from Tocris Bioscience (Ellisville, MO). 3-(4, 5-Dimethylthiazol-yl)-2,5-diphenyltetrazolium bromide (MTT), Dimethyl sulfoxide (DMSO) and verapamil were obtained from Sigma-Aldrich Co. (St. Louis, MO).
6
Protocols for Mast Cell Differentiation and Inhibition
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RBL-2H3 cells, obtained from ATCC (Manassas, VA, USA), were grown as monolayer cultures in minimal essential medium Eagles (MEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin at 37°C and 5% CO2. Depletion of the kinesin-1 heavy chain subunit, Kif5b, in RBL-2H3 cells was by RNA interference (RNAi) using MISSION lentiviral shRNAs (Sigma) from the mouse TRC1 library (TRC_ID: TRCN0000091[xxx], [xxx] = 479, 480 or 481). Mouse sequences were either identical to rat or contained one miss-match. Murine bone marrow-derived mast cells (BMMCs) were isolated as previously described [15 (link)]. BMMCs were cultured in RPMI media supplemented with 10% heat-inactivated FBS, non-essential amino acids (Gibco) and sodium pyruvate at 37°C and 5% CO2. Growth for four weeks in the presence of 20 ng/ml interleukin-3 was used to induce differentiation into mast cells. Maturation of mast cells was confirmed by FcεRI expression via flow cytometry. To test the effects of drugs, cells were pre-incubated for 30 min in 1 μM nocodazole (Sigma), 10 μM colchicine (Tocris) and 10 μM paclitaxel (Sigma), which target microtubules; and 100 μM kinesore (Tocris), which targets the microtubule motor kinesin-1. Drugs were dissolved in DMSO at 10 mM except kinesore which was dissolved at 20 mM; vehicle controls used DMSO at 0.5% (v/v).
7
Characterization of Neuromodulatory Compounds
8
Macrophage Activation and Cytoskeleton Regulation
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NIH-3T3 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Gibco) with 10% fetal bovine serum (FBS, Gibco) and 100 U/ml penicillin-streptomycin (pen-strep, Gibco). Bone marrow-derived macrophages were obtained from primary bone marrow cells cultured for 7 days in DMEM supplemented with 20% L929 culture supernatant, 10% FBS, and 100 U/ml pen-strep. We tried to use minimum mice to obtain bone marrow cells and protocol was approved by the Institutional Animal Care and Use Committee of the Yonsei Laboratory Animal Research Center at Yonsei University.
For OASL1 overexpression, plasmids were mixed with Lipofectamine 2000 (Invitrogen) and incuated at 37°C in 5% CO2 for 24 h. For stimulation, cells were treated with 10 μg/ml poly(I:C) (GE Healthcare), 100 ng/ml LPS (InvivoGen), 100 ng/ml Pam3CSK4 (InvivoGen), 10 μg/ml poly(dA:dT) (InvivoGen), and 1 μg/ml 5′ppp-dsRNA (InvivoGen) for the indicated times. For cytoskeleton inhibition experiments, cells were pre-treated with 10 ng/ml cytochalasin D (Sigma) and 100 μM colchicine (Tocris) for 30 min before poly(I:C) stimulation. To prevent autophagy assembly, cells were treated with 100 nM Bafilomycin A (Sigma), 5 mM 3-MA (3-methyladenine) (Sigma), and 50 μM CQ (chloroquine di-phosphate salt) (Sigma) along with poly(I:C).
For OASL1 overexpression, plasmids were mixed with Lipofectamine 2000 (Invitrogen) and incuated at 37°C in 5% CO2 for 24 h. For stimulation, cells were treated with 10 μg/ml poly(I:C) (GE Healthcare), 100 ng/ml LPS (InvivoGen), 100 ng/ml Pam3CSK4 (InvivoGen), 10 μg/ml poly(dA:dT) (InvivoGen), and 1 μg/ml 5′ppp-dsRNA (InvivoGen) for the indicated times. For cytoskeleton inhibition experiments, cells were pre-treated with 10 ng/ml cytochalasin D (Sigma) and 100 μM colchicine (Tocris) for 30 min before poly(I:C) stimulation. To prevent autophagy assembly, cells were treated with 100 nM Bafilomycin A (Sigma), 5 mM 3-MA (3-methyladenine) (Sigma), and 50 μM CQ (chloroquine di-phosphate salt) (Sigma) along with poly(I:C).
9
Colchicine Dorsal Hippocampus Injection
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We delivered 1 μL of 6 mg/mL colchicine (Tocris, Cat#
13–641-G) into the dorsal hippocampus at the following
coordinates AP −1.8, ML +2.5, DV −1.7 (in mm from Bregma)
in order to block axonal transport and increase the peptide amount in
cell bodies. Animals were perfused two days later and processed for
immunohistochemistry.
13–641-G) into the dorsal hippocampus at the following
coordinates AP −1.8, ML +2.5, DV −1.7 (in mm from Bregma)
in order to block axonal transport and increase the peptide amount in
cell bodies. Animals were perfused two days later and processed for
immunohistochemistry.
10
Cytotoxicity Screening of Anti-Cancer Drugs
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