Home >
Community >
How can I make the stock solution of AHL (3-oxo c12 HSL) in the...
Upvote
23
Downvote
+ Stock solution
+ Acetic acid
+ Ethyl acetate
Posted by
Loveleen Bajaj
How can I make the stock solution of AHL (3-oxo c12 HSL) in the...
Hello Vivek, now I am tryin to find the best protocol to make the aqueous stock solution of AHL (3-oxo c12 HSL), if you found the appropriate protocol, can you share it with me? Thank you.
Hello Vivek, now I am tryin to find the best protocol to make the aqueous stock solution of AHL (3-oxo c12 HSL), if you found the appropriate protocol, can you share it with me? Thank you.
Hello Vivek. My partner worked with the same AHL, and instead of glacial acetic acid, we used 0,05% formic acid with ethyl acetate to prepare our stocks of all AHLs that we use. We had the same situation (white precipitation). Once we prepare the stocks, we put at -20°C, then before of dissolving with the broth, we just resuspend in a vortex. We didn't notice a precipitation in our test, probably because the volume we used was minimal. Still it would be great to know if there is a properly way to dissolve this particular AHL since it is hydrophobic. I found this, hope it helps... We find that AHLs are best maintained as stock solutions in ethyl acetate acidified with glacial acetic acid (1 ml of acetic acid per 1 L of ethyl acetate). We store them at −20°C in glass vials. If borosilicate vials are used for this purpose, prior to adding AHL solutions, glass vials are first treated with 0.2 N HCl, and then rinsed extensively with deionized water. Teflon-lined caps should be used for capping vials containing organic solutions of AHLs. As needed for experiments, serial dilution of 3-oxo-C12-HSL can be prepared in other organic solvents (including ethanol, methanol and acetonitrile), however stability of the compound in these solutions is not known. 3-oxo-C12-HSL is not stable in aqueous solutions where it converts into tetramic acid, an antibiotic (22). At neutral or basic pH, AHLs undergo spontaneous lactonolysis (23). From Chapter 9 "Luminescent Reporters and Their Applications for the Characterization of Signals and Signal-Mimics that Alter LasR-Mediated Quorum Sensing" - Quorum Sensing: Methods and Protocols http://link.springer.com/protocol/10.1007%2F978-1-60761-971-0_9 Best regards.
Hello Vivek. My partner worked with the same AHL, and instead of glacial acetic acid, we used 0,05% formic acid with ethyl acetate to prepare our stocks of all AHLs that we use. We had the same situation (white precipitation). Once we prepare the stocks, we put at -20°C, then before of dissolving with the broth, we just resuspend in a vortex. We didn't notice a precipitation in our test, probably because the volume we used was minimal. Still it would be great to know if there is a properly way to dissolve this particular AHL since it is hydrophobic. I found this, hope it helps... We find that AHLs are best maintained as stock solutions in ethyl acetate acidified with glacial acetic acid (1 ml of acetic acid per 1 L of ethyl acetate). We store them at −20°C in glass vials. If borosilicate vials are used for this purpose, prior to adding AHL solutions, glass vials are first treated with 0.2 N HCl, and then rinsed extensively with deionized water. Teflon-lined caps should be used for capping vials containing organic solutions of AHLs. As needed for experiments, serial dilution of 3-oxo-C12-HSL can be prepared in other organic solvents (including ethanol, methanol and acetonitrile), however stability of the compound in these solutions is not known. 3-oxo-C12-HSL is not stable in aqueous solutions where it converts into tetramic acid, an antibiotic (22). At neutral or basic pH, AHLs undergo spontaneous lactonolysis (23). From Chapter 9 "Luminescent Reporters and Their Applications for the Characterization of Signals and Signal-Mimics that Alter LasR-Mediated Quorum Sensing" - Quorum Sensing: Methods and Protocols http://link.springer.com/protocol/10.1007%2F978-1-60761-971-0_9 Best regards.
Hello Vivek, now I am tryin to find the best protocol to make the aqueous stock solution of AHL (3-oxo c12 HSL), if you found the appropriate protocol, can you share it with me?
Thank you.
Hello Vivek, now I am tryin to find the best protocol to make the aqueous stock solution of AHL (3-oxo c12 HSL), if you found the appropriate protocol, can you share it with me?
Thank you.
More
VOTE
Hello Vivek.
My partner worked with the same AHL, and instead of glacial acetic acid, we used 0,05% formic acid with ethyl acetate to prepare our stocks of all AHLs that we use. We had the same situation (white precipitation). Once we prepare the stocks, we put at -20°C, then before of dissolving with the broth, we just resuspend in a vortex. We didn't notice a precipitation in our test, probably because the volume we used was minimal. Still it would be great to know if there is a properly way to dissolve this particular AHL since it is hydrophobic.
I found this, hope it helps...
We find that AHLs are best maintained as stock solutions in
ethyl acetate acidified with glacial acetic acid (1 ml of acetic
acid per 1 L of ethyl acetate). We store them at −20°C in glass
vials. If borosilicate vials are used for this purpose, prior to
adding AHL solutions, glass vials are first treated with 0.2 N
HCl, and then rinsed extensively with deionized water.
Teflon-lined caps should be used for capping vials containing
organic solutions of AHLs. As needed for experiments, serial
dilution of 3-oxo-C12-HSL can be prepared in other organic
solvents (including ethanol, methanol and acetonitrile), however
stability of the compound in these solutions is not known.
3-oxo-C12-HSL is not stable in aqueous solutions where it
converts into tetramic acid, an antibiotic (22). At neutral or
basic pH, AHLs undergo spontaneous lactonolysis (23).
From Chapter 9 "Luminescent Reporters and Their Applications for the Characterization of Signals and Signal-Mimics that Alter LasR-Mediated Quorum Sensing" - Quorum Sensing: Methods and Protocols
http://link.springer.com/protocol/10.1007%2F978-1-60761-971-0_9
Best regards.
Hello Vivek.
My partner worked with the same AHL, and instead of glacial acetic acid, we used 0,05% formic acid with ethyl acetate to prepare our stocks of all AHLs that we use. We had the same situation (white precipitation). Once we prepare the stocks, we put at -20°C, then before of dissolving with the broth, we just resuspend in a vortex. We didn't notice a precipitation in our test, probably because the volume we used was minimal. Still it would be great to know if there is a properly way to dissolve this particular AHL since it is hydrophobic.
I found this, hope it helps...
We find that AHLs are best maintained as stock solutions in
ethyl acetate acidified with glacial acetic acid (1 ml of acetic
acid per 1 L of ethyl acetate). We store them at −20°C in glass
vials. If borosilicate vials are used for this purpose, prior to
adding AHL solutions, glass vials are first treated with 0.2 N
HCl, and then rinsed extensively with deionized water.
Teflon-lined caps should be used for capping vials containing
organic solutions of AHLs. As needed for experiments, serial
dilution of 3-oxo-C12-HSL can be prepared in other organic
solvents (including ethanol, methanol and acetonitrile), however
stability of the compound in these solutions is not known.
3-oxo-C12-HSL is not stable in aqueous solutions where it
converts into tetramic acid, an antibiotic (22). At neutral or
basic pH, AHLs undergo spontaneous lactonolysis (23).
From Chapter 9 "Luminescent Reporters and Their Applications for the Characterization of Signals and Signal-Mimics that Alter LasR-Mediated Quorum Sensing" - Quorum Sensing: Methods and Protocols
http://link.springer.com/protocol/10.1007%2F978-1-60761-971-0_9
Best regards.
More
VOTE
Hi , you can see this publication
https://www.researchgate.net/publication/11162037_N-Acylhomoserine_Lactones_Undergo_Lactonolysis_in_a_pH-_Temperature-_and_Acyl_Chain_Length-Dependent_Manner_during_Growth_of_Yersinia_pseudotuberculosis_and_Pseudomonas_aeruginosa
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC128322/
Article N-Acylhomoserine Lactones Undergo Lactonolysis in a pH-, Tem...
Hi , you can see this publication
https://www.researchgate.net/publication/11162037_N-Acylhomoserine_Lactones_Undergo_Lactonolysis_in_a_pH-_Temperature-_and_Acyl_Chain_Length-Dependent_Manner_during_Growth_of_Yersinia_pseudotuberculosis_and_Pseudomonas_aeruginosa
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC128322/
Article N-Acylhomoserine Lactones Undergo Lactonolysis in a pH-, Tem...
More
VOTE
Hello Vivek, I would also request you to share the protocol for preparing the stock solution of AHL. Thank you
Hello Vivek, I would also request you to share the protocol for preparing the stock solution of AHL. Thank you
More
VOTE