RPMI-1640 and DMEM medium and foetal bovine serum (FBS) were obtained from GIBCO (Grand Island, NY). Trizol, PrimeScript RT Master Mix and SYBR green PCR master mix were purchased from Takara (Shiga, Japan). Antibodies against LC-3I/II, p62, VPS34, and HDAC6 were purchased from Cell Signaling Technology (Danvers, USA), antibodies against GFP, GRP78 and acetyl-lysine were from Abcam (Cambridge, UK), and anitbodies against β-tubulin, β-actin and GAPDH were from Abmart (Shanghai, China). The anti-CD63 antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, USA), and the anti-FITC-, -TRITC-, -Cy5- and -HRP-conjugated secondary antibodies as well as ER-Tracker and Lyso-Tracker were obtained from Invitrogen (Carlsbad, USA). Sodium butyrate, vorinostat (SAHA), tubastatin A and 3-methyladenine were obtained from Cayman Chemical (MI, USA). Anti-FLAG M2 mAb and anti-FLAG M2 agarose were obtained from Sigma (St. Louis, USA).
Sodium butyrate
Manufactured by Cayman Chemical
Sourced in United States
Sodium butyrate is a chemical compound used in various laboratory applications. It serves as a source of the butyrate anion, which plays a role in cellular processes. The product is available in a powdered form.
Automatically generated - may contain errors
Lab products found in correlation
Pmd2 g envelope plasmid, by Addgene (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Er tracker, by Thermo Fisher Scientific (1 mentions)
Anti cd63 antibody, by Santa Cruz Biotechnology (1 mentions)
3 methyladenine, by Cayman Chemical (1 mentions)
Anti flag m2 mab, by Merck Group (1 mentions)
Sybr green pcr master mix, by Takara Bio (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Anti flag m2 agarose, by Merck Group (1 mentions)
Lysotracker, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Primescript rt master mix, by Takara Bio (1 mentions)
Trizol, by Takara Bio (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Abmart (1 mentions)
Pspax2 packaging plasmid, by Addgene (1 mentions)
3 protocols using sodium butyrate
1
Autophagy regulation mechanisms
2
Lentiviral Particle Production and Transduction
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For the production of lentiviral particles, we used the second-generation lentiviral system according to Addgene instructions, using psPAX2 packaging plasmid and pMD2.G envelope plasmid (Addgene, Cambridge, MA). Co-transfection of the lentiviral packaging/envelope plasmids and transfer vector into the HEK239FT (2 × 107 cells/transfection) was performed using Lipofectamine 3000 (Thermo Fisher Scientific) scaled according to the manufacturer’s recommendations. After overnight incubation, sodium butyrate (Cayman Chemical) was added at a final concentration of 10 mM to increase viral titer. The lentiviral particles in the supernatant were collected at 48–72 h and used to transduce cells. GBM neurospheres (1.5 × 105 cells) were seeded in a 6-well cell culture plate and infected overnight with a lentiviral medium containing viral particles and polybrene (1 µg/mL), supplemented with the appropriate medium. On the following morning, cells were pelleted by centrifugation and resuspended in a fresh neurosphere medium. A list of lentiviral constructs used can be found in Supplementary Table 5 .
3
Lentiviral Particle Production and Transduction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the production of lentiviral particles, we used the 2 nd -generation lentiviral system according to Addgene instructions, using psPAX2 packaging plasmid and pMD2.G envelope plasmid (Addgene, Cambridge, MA). Co-transfection of the lentiviral packaging/envelope plasmids and transfer vector into the HEK239FT (2x10 7 cells/transfection) was performed using Lipofectamine 3000 (ThermoFisher Scienti c) scaled according to manufacturer recommendations. After overnight incubation, sodium butyrate (Cayman Chemical) was added at nal concentration of 10mM to increase viral titer. The lentiviral particles in supernatant were collected at 48-72 h and used to transduce cells. GBM neurospheres (1.5 × 10 5 cells) were seeded in a 6-well cell culture plate and infected overnight with lentiviral medium containing viral particles and polybrene (1 µg/mL), supplemented with appropriate medium. On the following morning, cells were pelleted by centrifugation and resuspended in fresh neurosphere medium. List of lentiviral constructs used can be found in table S5.
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