How do you dissolve 3-Ma?
3-MA is soluble in water, but only at a relatively high temperature (>40C for 10mM solution). I would suggest preparing a stock solution of 100mM concentration in water by warming to around 60C, then aliquoting and storing at -20C. Each aliquot will need to be redissolved at around 40C when required.
How 3-MA inhibit autophagy?
3-Methyladenine (3- MA) is an inhibitor of phosphatidylinositol 3-kinases (PI3K). PI3K plays an important role in many biological processes, including controlling the activation of mTOR, a key regulator of autophagy. 3-MA inhibits autophagy by blocking autophagosome formation via the inhibition of class III PI3K [1].
How does LY294002 inhibit PI3K?
The PI3K inhibitor LY294002 prevents induction of p53 by DNA damage. To explore the contribution of particular signal transduction cascades to the effect of serum growth factors on p53 induction, cells were exposed to CisPt in the presence of specific chemical inhibitors.
How does wortmannin inhibit PI3K?
Wortmannin is a potent irreversible inhibitor of the PI3K pathway that binds the ATP-binding site of p110 kinase domain, preventing the conversion to PIP3 (Powis et al., 1994).
What is Bafilomycin used for?
Bafilomycin A1 is widely used as an inhibitor of autophagosome-lysosome fusion in vitro to determine the activity of autophagic flux. Bafilomycin A1 is a macrolide that targets the V-ATPase ATP6V0C/V0 subunit c, thus inhibiting lysosomal acidification by preventing the passage of protons into the lysosomal lumen.
What is an autophagy inhibitor?
Autophagy inhibitors, such as chloroquine and hydroxychloroquine, have already been clinically approved, promoting drug combination treatment by targeting autophagic pathways as a means of discovering and developing more novel and more effective cancer therapeutic approaches.
What is wortmannin used for?
Wortmannin is a commonly used cell biology reagent that has been used previously in research to inhibit DNA repair, receptor-mediated endocytosis and cell proliferation.
How do you dissolve Bafilomycin?
Bafilomycin A1 is supplied as a crystalline solid. A stock solution may be made by dissolving the bafilomycin A1 in the solvent of choice, which should be purged with an inert gas. Bafilomycin A1 is soluble in organic solvents such as DMSO and methanol.
How does Bafilomycin A1 inhibit autophagy?
We found that bafilomycin A1 blocks autophagosome-lysosome fusion and lysosome acidification in Drosophila fat body cells, as it does in vitro in mammalian cell culture. However, our genetic results demonstrate that V-ATPase-dependent acidification is not a prerequisite for vesicle fusion.
What kind of liquid is DMSO miscible with?
DMSO also dissolves many inorganic salts, particularly transition metals nitrates, cyanides and dichromates. DMSO is miscible with water and most organic liquids. The contents of this page summarize solubility characteristics for DMSO.
Why is the solubility of DMSO compounds lower?
Solubility of compounds in wet-DMSO is much lower than in dry DMSO. A small amount of water in the DMSO can decrease solubility dramatically, due to the non-ideal properties of DMSO-water mixtures. Cavity formation during dissolution is more difficult in wet DMSO than in dry DMSO or in pure water (Oldenburg et al., 2005).
Which is the best concentration to store DMSO?
When compound stock solutions are stored at lower concentrations, solubility in DMSO is less of a problem. The optimal storage concentrations were found to be 2-5 mM, which maximizes solubility and minimizes precipitation in DMSO (Popa-Burke et al., 2004; Schopfer et al., 2005).
How long does it take DMSO to absorb 10% water?
DMSO is very hygroscopic and will absorb up to 10% water in as little as 6 hours under laboratory conditions (Fillers, 2004). In fact, it is technically difficult to limit uptake of water into DMSO. A few companies deliberately introduce about 10% water into the DMSO and store stock solutions in a cool but nonsolid matrix (Schopfer etal., 2005).