Sneha Singh well ,the best solvent indeed was plasma itself, i.e. re-suspending the concentrate in plasma to reach the desired concentration. but hepes/tris/PBS and even citrate will also work so long as to keep pH around 7 and using physical (not chemical) precipitation method to purify fibrinogen. chemically precipitated fg will solve at a much slower rate. perhaps due to binding of small ligand molecules used in chemical precipitation methods which needs to be removed before re-suspending fg in solvent.
Sneha Singh well ,the best solvent indeed was plasma itself, i.e. re-suspending the concentrate in plasma to reach the desired concentration. but hepes/tris/PBS and even citrate will also work so long as to keep pH around 7 and using physical (not chemical) precipitation method to purify fibrinogen. chemically precipitated fg will solve at a much slower rate. perhaps due to binding of small ligand molecules used in chemical precipitation methods which needs to be removed before re-suspending fg in solvent.
Thanks for time spent. But Tris buffer is cyrotoxic to my knowledge and i want to use fibrinogen for cell seeding scaffold fabrication. Any idea avout sodium citeate suitability?
Thanks for time spent. But Tris buffer is cyrotoxic to my knowledge and i want to use fibrinogen for cell seeding scaffold fabrication. Any idea avout sodium citeate suitability?
No, I mean 4 centigrade degrees not 40! put it on rotator in the cold room where the temperature is +4.
No, I mean 4 centigrade degrees not 40! put it on rotator in the cold room where the temperature is +4.
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Mohamadhasan Ansarizadeh ok. tnx for the contribution
Mohamadhasan Ansarizadeh ok. tnx for the contribution
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Ok, I discussed with someone about your issue.
Hey suggested that search some papers in which fibrinogen treated with Tris-HCl buffer.
Ok, I discussed with someone about your issue.
Hey suggested that search some papers in which fibrinogen treated with Tris-HCl buffer.
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Sodium citrate. Sorry for misspell
Sodium citrate. Sorry for misspell
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I just add PBS to reach 25 mg/ml concentration and then rock it overnight at 4OC. Never agitate it by hand
I just add PBS to reach 25 mg/ml concentration and then rock it overnight at 4OC. Never agitate it by hand
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Hi Rooney
Did it work for you? I am attempting to purify fgn, which buffer has worked for you finally. This will give me a direction too.
Thanks
Hi Rooney
Did it work for you? I am attempting to purify fgn, which buffer has worked for you finally. This will give me a direction too.
Thanks
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Sneha Singh well ,the best solvent indeed was plasma itself, i.e. re-suspending the concentrate in plasma to reach the desired concentration. but hepes/tris/PBS and even citrate will also work so long as to keep pH around 7 and using physical (not chemical) precipitation method to purify fibrinogen. chemically precipitated fg will solve at a much slower rate. perhaps due to binding of small ligand molecules used in chemical precipitation methods which needs to be removed before re-suspending fg in solvent.
Sneha Singh well ,the best solvent indeed was plasma itself, i.e. re-suspending the concentrate in plasma to reach the desired concentration. but hepes/tris/PBS and even citrate will also work so long as to keep pH around 7 and using physical (not chemical) precipitation method to purify fibrinogen. chemically precipitated fg will solve at a much slower rate. perhaps due to binding of small ligand molecules used in chemical precipitation methods which needs to be removed before re-suspending fg in solvent.
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@prashant pandey
Thanks for time spent. But Tris buffer is cyrotoxic to my knowledge and i want to use fibrinogen for cell seeding scaffold fabrication. Any idea avout sodium citeate suitability?
@prashant pandey
Thanks for time spent. But Tris buffer is cyrotoxic to my knowledge and i want to use fibrinogen for cell seeding scaffold fabrication. Any idea avout sodium citeate suitability?
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Mohamadhasan Ansarizadeh Thanks for your contribution. U mean going beyond 37 C and using an automatic shaker overnight should resolve the issue?
Mohamadhasan Ansarizadeh Thanks for your contribution. U mean going beyond 37 C and using an automatic shaker overnight should resolve the issue?
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Sorry for the misspell
I meant Roozbeh not Rooney
Sorry for the misspell
I meant Roozbeh not Rooney
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