For cell spreading studies, the following concentrations were used: 50 µM blebbistatin, 30 µM Y-27632, 1 µM Cytochalasin D (all from Wako, Japan) and 2 mM GRGDS (Peptide institute, Japan). Each of these reagents was added 2 h prior to analysis. For lipid raft and pFAK studies, 25 µM methyl-β-cyclodextrin (MβCD, Santa Cruz Biotechnology) was added when seeding the cells. MβCD was incubated for 2 days prior to analysis. For differentiation studies, 25 µM MβCD and 4 µM FAK inhibitor 14 (Sigma-Aldrich) were used. MβCD was incubated until day 14 and was exchanged every 3 d. FAK inhibitor 14 was incubated until day 2 and then was exchanged for fresh growth medium.
Methyl β cyclodextrin mβcd
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Methyl-β-cyclodextrin (MβCD) is a cyclic oligosaccharide derivative used in various laboratory applications. It functions as a complexing agent, capable of forming inclusion complexes with a wide range of compounds.
Automatically generated - may contain errors
Lab products found in correlation
Blebbistatin, by Fujifilm (1 mentions)
Fak inhibitor 14, by Merck Group (1 mentions)
Y 27632, by Fujifilm (1 mentions)
Cytochalasin d, by Fujifilm (1 mentions)
Anti chop, by Cell Signaling Technology (1 mentions)
Anti vdac, by Cell Signaling Technology (1 mentions)
Anti bip, by Cell Signaling Technology (1 mentions)
Anti ire1α, by Cell Signaling Technology (1 mentions)
Z lehd fmk, by R&D Systems (1 mentions)
Anti atf6α, by Santa Cruz Biotechnology (1 mentions)
Anti cox 2, by Santa Cruz Biotechnology (1 mentions)
Pan caspase inhibitor z vad fmk, by R&D Systems (1 mentions)
N acetyl l cysteine nac, by Merck Group (1 mentions)
Anti caspase 12, by Cell Signaling Technology (1 mentions)
Anti p ire1α, by Thermo Fisher Scientific (1 mentions)
Anti cytochrome c, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit igg, by Abcam (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti caspase 8, by Cell Signaling Technology (1 mentions)
Anti caspase 9, by Cell Signaling Technology (1 mentions)
3 protocols using methyl β cyclodextrin mβcd
1
Cell Spreading and Differentiation Protocols
2
Exploring Apoptosis Pathways in Cells
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Synthetic N-hexanoyl-DL-homoserine lactone (C6-HSL), N-decanoyl-DL-homoserine lactone (C10-HSL), N-(3-hydroxy dodecanoyl)-DL-homoserine lactone (OH-dDHL), tunicamycin, thapsigargin, hydrogen peroxide 30% (wt/vol) (H2O2), N-Acetyl-l-cysteine (NAC), and staurosporine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies with anti-caspase-3, anti-caspase-8, anti-cytochrome c, anti-PARP, anti-β-actin, anti-caspase-9, anti-VDAC, anti-CHOP, anti- caspase-12, anti-p-eIF2α, anti-IRE1α, and anti-BIP were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-p-IRE1α was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Anti-ATF 6α, anti-COX-2, and methyl-β-cyclodextrin (MβCD) were purchased from Santa Cruz Biotechnology (Paso Robles, CA, USA). Horseradish peroxidase-conjugated goat anti-rabbit IgG and goat anti-mouse IgG were purchased from Abcam (Cambridge, MA, USA). The pan-caspase inhibitor (Z-VAD-FMK), caspase-3 inhibitor (Z-DEVE-FMK), and caspase-9 inhibitor (Z-LEHD-FMK) were purchased from R&D Systems (Minneapolis, MN, USA). The paraoxonase 2 (PON2) inhibitor ((1,2,4)trizolo(4,3–a)quinolones; TQ416) was purchased from Maybridge (Cornwall, UK).
3
Lipid Raft Disruption and Antibody Analysis
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Methyl-β-cyclodextr in (MβCD) and anti-neutrophil myeloperoxidase [MPO; mouse anti-goat polyclonal IgG (H+L)] antibodies were obtained from Santa Cruz Biotechnology, Inc. (1:100, cat. no. sc-16129, Dallas, TX, USA), along with lipid-raft disruptor filipin. Alexa Fluor 488-cholera toxin subunit B (CTXB) and Alexa Fluor 594-labeled chicken anti-Goat IgG (H+L) secondary antibody were purchased from Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA). Anti-flotillin-1 antibody was purchased from BD Biosciences (1:1000, cat no. 61802; Franklin Lakes, NJ, USA). Polyclonal anti-ERp29 rabbit anti-mouse antibody was obtained from Abcam (1:3000, ab11420; Cambridge, MA, USA). Optiprep was obtained from Axis-Shield, Inc. (Norton, MA, USA). High glucose Dulbecco's modified Eagle's medium (DMEM) was purchased from GE Healthcare Life Sciences (Logan, UT, USA). An Enhanced Chemiluminescence (ECL) kit was obtained from PerkinElmer Inc. (Waltham, MA, USA). Human LDL was purchased from ProSpec-Tany TechnoGene, Ltd. (Ness Ziona, Israel).
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