THE PURPOSE OF USING GW4869 IS TO INHIBIT EXOSOME RELEASE. BOTH 5 & 10 are working in cells. It may vary upon the cells u are using. so perform a mtt to know the toxicity and then for effective concentration in ur cell line refer literature
THE PURPOSE OF USING GW4869 IS TO INHIBIT EXOSOME RELEASE. BOTH 5 & 10 are working in cells. It may vary upon the cells u are using. so perform a mtt to know the toxicity and then for effective concentration in ur cell line refer literature
GW4869 prepared at conc of 1mM is not turbid or opaque and will be clear. Either U can dilute from the stock 5mM (looks pale white or turbid) to 1mM (clear solution). U can see a clear solution in the conc of 1mM.
GW4869 prepared at conc of 1mM is not turbid or opaque and will be clear. Either U can dilute from the stock 5mM (looks pale white or turbid) to 1mM (clear solution). U can see a clear solution in the conc of 1mM.
I am also having the same problem. I found this publication (http://www.jimmunol.org/content/185/11/6960.full) in which they generate a 7.5 mM Stock solution with DMSO and before adding it to cell culture supernatant they supplement 1/20 volume of 5% methanesulfonic acid to help solubilize.
I am also having the same problem. I found this publication (http://www.jimmunol.org/content/185/11/6960.full) in which they generate a 7.5 mM Stock solution with DMSO and before adding it to cell culture supernatant they supplement 1/20 volume of 5% methanesulfonic acid to help solubilize.
Dear, you can dissolve GW4869 in DMSO as a suspension stock of 10 mM, then dilute it into working concentration (10-20 uM) by medium (e.g. DEME) with a clear solution. The compound from MCE has been verified.
Dear, you can dissolve GW4869 in DMSO as a suspension stock of 10 mM, then dilute it into working concentration (10-20 uM) by medium (e.g. DEME) with a clear solution. The compound from MCE has been verified.
Dear Guo-Chang Fan, thank you for your input. I dissolved mine in DMSO as reported on the Sigma website (0.2mg/ml) and then diluted (x10) in PBS to get the working concentrations I need. however, i am interested to know how you prepared the stock of 5mM since 0.2mg/ml correpsonds to 0.346mM
Dear Guo-Chang Fan, thank you for your input. I dissolved mine in DMSO as reported on the Sigma website (0.2mg/ml) and then diluted (x10) in PBS to get the working concentrations I need. however, i am interested to know how you prepared the stock of 5mM since 0.2mg/ml correpsonds to 0.346mM
I tried dissolving GW4869 in DMSO to get stock of 10mM, but it was turbid and I can see some particulate. Thus I tried to dilute the stock further and at 3mM the solution started getting clear. 1mM was the dilution when I can see a clear solution of GW4869 and I am using 10uM final concentration for my experiment which is working perfectly for me.
I tried dissolving GW4869 in DMSO to get stock of 10mM, but it was turbid and I can see some particulate. Thus I tried to dilute the stock further and at 3mM the solution started getting clear. 1mM was the dilution when I can see a clear solution of GW4869 and I am using 10uM final concentration for my experiment which is working perfectly for me.
Yvette Wilda Jyrwa I work on platelets; thus, my treatment is 10min 10uM Gw4869. For long-incubation, you should use 1uM final concentration, equivalent to IC 50 of the compound. I used DMSO to dissolve Gw4869 to a stock concentration of 1mM without any difficulty.
Yvette Wilda Jyrwa I work on platelets; thus, my treatment is 10min 10uM Gw4869. For long-incubation, you should use 1uM final concentration, equivalent to IC 50 of the compound. I used DMSO to dissolve Gw4869 to a stock concentration of 1mM without any difficulty.
Hi Ranjithkumar, What is the conc. of GW4869 that you are using in cells? Do you have a specific conc. which works in cells? what cells are you treating the GW4869? Do you do pre-treatment or combination treatment to look for its effect in cells?
Hi Ranjithkumar, What is the conc. of GW4869 that you are using in cells? Do you have a specific conc. which works in cells? what cells are you treating the GW4869? Do you do pre-treatment or combination treatment to look for its effect in cells?
Did GW4869 at 5 or 10 um work for your experiments? what outcomes are you measuring after treatment in epithelial cells?
Did GW4869 at 5 or 10 um work for your experiments? what outcomes are you measuring after treatment in epithelial cells?
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THE PURPOSE OF USING GW4869 IS TO INHIBIT EXOSOME RELEASE. BOTH 5 & 10 are working in cells. It may vary upon the cells u are using. so perform a mtt to know the toxicity and then for effective concentration in ur cell line refer literature
THE PURPOSE OF USING GW4869 IS TO INHIBIT EXOSOME RELEASE. BOTH 5 & 10 are working in cells. It may vary upon the cells u are using. so perform a mtt to know the toxicity and then for effective concentration in ur cell line refer literature
More
VOTE
GW4869 prepared at conc of 1mM is not turbid or opaque and will be clear.
Either U can dilute from the stock 5mM (looks pale white or turbid) to 1mM (clear solution).
U can see a clear solution in the conc of 1mM.
GW4869 prepared at conc of 1mM is not turbid or opaque and will be clear.
Either U can dilute from the stock 5mM (looks pale white or turbid) to 1mM (clear solution).
U can see a clear solution in the conc of 1mM.
More
VOTE
Am using epithelial cell line. Conc of GW4869 used was 5 and 10 uM. I co incubated GW4869 along with my treatment.
Am using epithelial cell line. Conc of GW4869 used was 5 and 10 uM. I co incubated GW4869 along with my treatment.
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VOTE
I am also having the same problem. I found this publication (http://www.jimmunol.org/content/185/11/6960.full) in which they generate a 7.5 mM Stock solution with DMSO and before adding it to cell culture supernatant they supplement 1/20 volume of 5% methanesulfonic acid to help solubilize.
I am also having the same problem. I found this publication (http://www.jimmunol.org/content/185/11/6960.full) in which they generate a 7.5 mM Stock solution with DMSO and before adding it to cell culture supernatant they supplement 1/20 volume of 5% methanesulfonic acid to help solubilize.
More
VOTE
Dear, you can dissolve GW4869 in DMSO as a suspension stock of 10 mM, then dilute it into working concentration (10-20 uM) by medium (e.g. DEME) with a clear solution. The compound from MCE has been verified.
Dear, you can dissolve GW4869 in DMSO as a suspension stock of 10 mM, then dilute it into working concentration (10-20 uM) by medium (e.g. DEME) with a clear solution. The compound from MCE has been verified.
More
VOTE
But the solubility of GW4869 in DMSO is 0.2 mg/mL, which is 0.346 mM. How did they get the stock solution of 5 mM in DMSO?
But the solubility of GW4869 in DMSO is 0.2 mg/mL, which is 0.346 mM. How did they get the stock solution of 5 mM in DMSO?
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Santosh Yadav For long do you treat your cells with GW4869? I'm in doubt about what procedure to follow. Please help me with the procedure.
Santosh Yadav For long do you treat your cells with GW4869? I'm in doubt about what procedure to follow. Please help me with the procedure.
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VOTE
Dear Guo-Chang Fan, thank you for your input. I dissolved mine in DMSO as reported on the Sigma website (0.2mg/ml) and then diluted (x10) in PBS to get the working concentrations I need. however, i am interested to know how you prepared the stock of 5mM since 0.2mg/ml correpsonds to 0.346mM
Dear Guo-Chang Fan, thank you for your input. I dissolved mine in DMSO as reported on the Sigma website (0.2mg/ml) and then diluted (x10) in PBS to get the working concentrations I need. however, i am interested to know how you prepared the stock of 5mM since 0.2mg/ml correpsonds to 0.346mM
More
VOTE
I tried dissolving GW4869 in DMSO to get stock of 10mM, but it was turbid and I can see some particulate. Thus I tried to dilute the stock further and at 3mM the solution started getting clear. 1mM was the dilution when I can see a clear solution of GW4869 and I am using 10uM final concentration for my experiment which is working perfectly for me.
I tried dissolving GW4869 in DMSO to get stock of 10mM, but it was turbid and I can see some particulate. Thus I tried to dilute the stock further and at 3mM the solution started getting clear. 1mM was the dilution when I can see a clear solution of GW4869 and I am using 10uM final concentration for my experiment which is working perfectly for me.
More
VOTE
Yvette Wilda Jyrwa I work on platelets; thus, my treatment is 10min 10uM Gw4869. For long-incubation, you should use 1uM final concentration, equivalent to IC 50 of the compound. I used DMSO to dissolve Gw4869 to a stock concentration of 1mM without any difficulty.
Yvette Wilda Jyrwa I work on platelets; thus, my treatment is 10min 10uM Gw4869. For long-incubation, you should use 1uM final concentration, equivalent to IC 50 of the compound. I used DMSO to dissolve Gw4869 to a stock concentration of 1mM without any difficulty.
More
VOTE
Hi Ranjithkumar, What is the conc. of GW4869 that you are using in cells? Do you have a specific conc. which works in cells? what cells are you treating the GW4869? Do you do pre-treatment or combination treatment to look for its effect in cells?
Hi Ranjithkumar, What is the conc. of GW4869 that you are using in cells? Do you have a specific conc. which works in cells? what cells are you treating the GW4869? Do you do pre-treatment or combination treatment to look for its effect in cells?
More
VOTE