Re: [ccp4bb] Solubility of Aluminum Fluoride

[Firdous Tarique]

Thanks everyone for their wonderful suggestions.

Best

F

On Wed, Jun 22, 2022 at 11:42 AM Patrick Loll  wrote:

> Kevin and Guillaume said it all quite well. I just want to stress one
> thing, namely that these solutions are metastable, and will eventually
> precipitate (hours to days).
>
> In the case of the beryllium salts, the precipitation is due to slow
> formation of metal hydroxide species. I have no direct experience with the
> aluminum variants, but since aluminum is notorious for forming goopy
> hydroxides, I would guess that the same issue pertains. So make the
> solution right before using, try not to use a pH too far above neutrality,
> and be very cautious about (for example) putting any of these solutions
> into an FPLC.
>
> Pat
>
> > On 22 Jun 2022, at 12:15 PM, Dr. Kevin M Jude 
> wrote:
> >
> > Sorry, I should have said ADPBeF3 (not ADPBeF4), and increase the NaF
> proportion for ADPAlF4. Which in practice means reducing the concentrations
> of everything else due to limited solubility of NaF.
> >
> > From: CCP4 bulletin board  on behalf of Dr.
> Kevin M Jude 
> > Date: Wednesday, June 22, 2022 at 9:10 AM
> > To: CCP4BB@JISCMAIL.AC.UK 
> > Subject: Re: [ccp4bb] Solubility of Aluminum Fluoride
> >
> > I posted the following to ccp4bb in 2017 in response to a similar
> question about ADPBeF4. For AlF4 you’ll increase the NaF stoichiometrically:
> >
> > reagent[stock] (mM)final concentration (mM)volume (µl)
>
> > BeCl2  100090  9
> > NaF750 450 60
> > ADP, pH 8  100 30  30
> > H2O1
> > total volume   100
> >
> > To prepare ADP:Be:F (1:3:15)
> > Prepare BeCl2 stock in the fume hood due to release of Cl2 gas
>
> > thoroughly mix BeCl2 and NaF
> > Add ADP
> > dilute with H2O to final volume
> >
> > This is done at room temperature and the reactions are fast. Careful
> mixing (and adjusting the pH of the ADP beforehand) are important because
> the BeCl2 stock will precipitate at neutral or elevated pH, and the BeFx
> solution is quite acidic.
> >
> >
> >
>
> ---
> Patrick J. Loll, Ph. D.  (he, him, his)
> Professor of Biochemistry & Molecular Biology
> Drexel University College of Medicine
> Room 10-102 New College Building
> 245 N. 15th St., Mailstop 497
> Philadelphia, PA  19102  USA
>
> (215) 762-7706
> pjl...@gmail.com
> pj...@drexel.edu
>
> 
>
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Re: [ccp4bb] Solubility of Aluminum Fluoride

[Patrick Loll]

Kevin and Guillaume said it all quite well. I just want to stress one thing, 
namely that these solutions are metastable, and will eventually precipitate 
(hours to days). 

In the case of the beryllium salts, the precipitation is due to slow formation 
of metal hydroxide species. I have no direct experience with the aluminum 
variants, but since aluminum is notorious for forming goopy hydroxides, I would 
guess that the same issue pertains. So make the solution right before using, 
try not to use a pH too far above neutrality, and be very cautious about (for 
example) putting any of these solutions into an FPLC. 

Pat

> On 22 Jun 2022, at 12:15 PM, Dr. Kevin M Jude  wrote:
> 
> Sorry, I should have said ADPBeF3 (not ADPBeF4), and increase the NaF 
> proportion for ADPAlF4. Which in practice means reducing the concentrations 
> of everything else due to limited solubility of NaF.
>  
> From: CCP4 bulletin board  on behalf of Dr. Kevin M 
> Jude 
> Date: Wednesday, June 22, 2022 at 9:10 AM
> To: CCP4BB@JISCMAIL.AC.UK 
> Subject: Re: [ccp4bb] Solubility of Aluminum Fluoride
> 
> I posted the following to ccp4bb in 2017 in response to a similar question 
> about ADPBeF4. For AlF4 you’ll increase the NaF stoichiometrically:
>  
> reagent[stock] (mM)final concentration (mM)volume (µl)
> BeCl2  100090  9
> NaF750 450 60
> ADP, pH 8  100 30  30
> H2O1
> total volume   100
> 
> To prepare ADP:Be:F (1:3:15)
> Prepare BeCl2 stock in the fume hood due to release of Cl2 gas
> 
> thoroughly mix BeCl2 and NaF
> Add ADP
> dilute with H2O to final volume
> 
> This is done at room temperature and the reactions are fast. Careful mixing 
> (and adjusting the pH of the ADP beforehand) are important because the BeCl2 
> stock will precipitate at neutral or elevated pH, and the BeFx solution is 
> quite acidic.
>  
>  
> 
---
Patrick J. Loll, Ph. D.  (he, him, his)
Professor of Biochemistry & Molecular Biology
Drexel University College of Medicine
Room 10-102 New College Building
245 N. 15th St., Mailstop 497
Philadelphia, PA  19102  USA

(215) 762-7706
pjl...@gmail.com
pj...@drexel.edu



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Re: [ccp4bb] Solubility of Aluminum Fluoride

[Dr. Kevin M Jude]

Sorry, I should have said ADPBeF3 (not ADPBeF4), and increase the NaF 
proportion for ADPAlF4. Which in practice means reducing the concentrations of 
everything else due to limited solubility of NaF.

From: CCP4 bulletin board  on behalf of Dr. Kevin M Jude 

Date: Wednesday, June 22, 2022 at 9:10 AM
To: CCP4BB@JISCMAIL.AC.UK 
Subject: Re: [ccp4bb] Solubility of Aluminum Fluoride
I posted the following to ccp4bb in 2017 in response to a similar question 
about ADPBeF4. For AlF4 you’ll increase the NaF stoichiometrically:

reagent[stock] (mM)final concentration (mM)volume (µl)
BeCl2  100090  9
NaF750 450 60
ADP, pH 8  100 30  30
H2O1
total volume   100

To prepare ADP:Be:F (1:3:15)
Prepare BeCl2 stock in the fume hood due to release of Cl2 gas
thoroughly mix BeCl2 and NaF
Add ADP
dilute with H2O to final volume
This is done at room temperature and the reactions are fast. Careful mixing 
(and adjusting the pH of the ADP beforehand) are important because the BeCl2 
stock will precipitate at neutral or elevated pH, and the BeFx solution is 
quite acidic.


From: CCP4 bulletin board  on behalf of Firdous Tarique 

Date: Tuesday, June 21, 2022 at 5:13 PM
To: CCP4BB@JISCMAIL.AC.UK 
Subject: Re: [ccp4bb] Solubility of Aluminum Fluoride
Thanks for your input. To avoid precipitation what about doing something like 
this? For example, as we know: AlCl3 + 3NaF = AlF3 + 3NaCl

Therefore, adding 100mM AlCl3 plus 300mM NaF to get 100mM of AlF3 solution, 
then directly using this 100 mM stock of AlF3 for the final dilution, say 1 or 
2 mM in the reaction mix ? Or it is recommended to use AlCl3 and NaF hoping to 
form AlF3 in the reaction mix.

Best

Faisal


On Tue, Jun 21, 2022 at 2:08 PM Guillaume Gaullier 
mailto:guillaume.gaull...@univ-tlse3.fr>> 
wrote:
Hi Firdous,

I have read a fair amount of papers about ATP-dependent chromatin remodelers in 
which ADP-BeF3 is often cited. It is made as follows (quoting from one of these 
papers): "ADP-BeF3 was prepared as a 10x stock (10 mM ADP, 30 mM BeSO4, 150 mM 
NaF, 10 mM MgCl2), freshly before use."

So the trick is to get all you need from a mixture of soluble compounds, 
instead of from only one poorly soluble salt. Not sure this will work the same 
with Al instead of Be, but it’s easy enough to try: use any known-soluble salt 
of Al instead of this BeSO4, and see if it works. NaF is extremely toxic, so 
use adequate protective measures (PPE, hood, etc.).

Good luck,

Guillaume




On 21 Jun 2022, at 19:38, Firdous Tarique 
mailto:kahkashantari...@gmail.com>> wrote:

Hi

Sorry for the off topic question but can anybody please suggest how to dissolve 
Aluminium Fluoride in water ? I know it has little solubility in water but have 
seen people using AF3 for their structural studies (ADP + AF3 as ATP analogue). 
Is there any other solvent where they have made AF3 solution or is there some 
trick to dissolve it in water?.

Thanks

Firdous



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Re: [ccp4bb] Solubility of Aluminum Fluoride

[Dr. Kevin M Jude]

I posted the following to ccp4bb in 2017 in response to a similar question 
about ADPBeF4. For AlF4 you’ll increase the NaF stoichiometrically:

reagent[stock] (mM)final concentration (mM)volume (µl)
BeCl2  100090  9
NaF750 450 60
ADP, pH 8  100 30  30
H2O1
total volume   100

To prepare ADP:Be:F (1:3:15)
Prepare BeCl2 stock in the fume hood due to release of Cl2 gas
thoroughly mix BeCl2 and NaF
Add ADP
dilute with H2O to final volume
This is done at room temperature and the reactions are fast. Careful mixing 
(and adjusting the pH of the ADP beforehand) are important because the BeCl2 
stock will precipitate at neutral or elevated pH, and the BeFx solution is 
quite acidic.


From: CCP4 bulletin board  on behalf of Firdous Tarique 

Date: Tuesday, June 21, 2022 at 5:13 PM
To: CCP4BB@JISCMAIL.AC.UK 
Subject: Re: [ccp4bb] Solubility of Aluminum Fluoride
Thanks for your input. To avoid precipitation what about doing something like 
this? For example, as we know: AlCl3 + 3NaF = AlF3 + 3NaCl

Therefore, adding 100mM AlCl3 plus 300mM NaF to get 100mM of AlF3 solution, 
then directly using this 100 mM stock of AlF3 for the final dilution, say 1 or 
2 mM in the reaction mix ? Or it is recommended to use AlCl3 and NaF hoping to 
form AlF3 in the reaction mix.

Best

Faisal


On Tue, Jun 21, 2022 at 2:08 PM Guillaume Gaullier 
mailto:guillaume.gaull...@univ-tlse3.fr>> 
wrote:
Hi Firdous,

I have read a fair amount of papers about ATP-dependent chromatin remodelers in 
which ADP-BeF3 is often cited. It is made as follows (quoting from one of these 
papers): "ADP-BeF3 was prepared as a 10x stock (10 mM ADP, 30 mM BeSO4, 150 mM 
NaF, 10 mM MgCl2), freshly before use."

So the trick is to get all you need from a mixture of soluble compounds, 
instead of from only one poorly soluble salt. Not sure this will work the same 
with Al instead of Be, but it’s easy enough to try: use any known-soluble salt 
of Al instead of this BeSO4, and see if it works. NaF is extremely toxic, so 
use adequate protective measures (PPE, hood, etc.).

Good luck,

Guillaume



On 21 Jun 2022, at 19:38, Firdous Tarique 
mailto:kahkashantari...@gmail.com>> wrote:

Hi

Sorry for the off topic question but can anybody please suggest how to dissolve 
Aluminium Fluoride in water ? I know it has little solubility in water but have 
seen people using AF3 for their structural studies (ADP + AF3 as ATP analogue). 
Is there any other solvent where they have made AF3 solution or is there some 
trick to dissolve it in water?.

Thanks

Firdous



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Re: [ccp4bb] Solubility of Aluminum Fluoride

[Guillaume Gaullier]

I have never tried varying the order of mixing, but it seems the correct 
ADP-BeF3 complex was formed when mixing all three solutions just before use.

Guillaume


> On 22 Jun 2022, at 02:12, Firdous Tarique  wrote:
> 
> Thanks for your input. To avoid precipitation what about doing something like 
> this? For example, as we know: AlCl3 + 3NaF = AlF3 + 3NaCl
> 
> Therefore, adding 100mM AlCl3 plus 300mM NaF to get 100mM of AlF3 solution, 
> then directly using this 100 mM stock of AlF3 for the final dilution, say 1 
> or 2 mM in the reaction mix ? Or it is recommended to use AlCl3 and NaF 
> hoping to form AlF3 in the reaction mix.
> 
> Best
> 
> Faisal
> 
> 
> On Tue, Jun 21, 2022 at 2:08 PM Guillaume Gaullier 
> mailto:guillaume.gaull...@univ-tlse3.fr>> 
> wrote:
> Hi Firdous,
> 
> I have read a fair amount of papers about ATP-dependent chromatin remodelers 
> in which ADP-BeF3 is often cited. It is made as follows (quoting from one of 
> these papers): "ADP-BeF3 was prepared as a 10x stock (10 mM ADP, 30 mM BeSO4, 
> 150 mM NaF, 10 mM MgCl2), freshly before use."
> 
> So the trick is to get all you need from a mixture of soluble compounds, 
> instead of from only one poorly soluble salt. Not sure this will work the 
> same with Al instead of Be, but it’s easy enough to try: use any 
> known-soluble salt of Al instead of this BeSO4, and see if it works. NaF is 
> extremely toxic, so use adequate protective measures (PPE, hood, etc.).
> 
> Good luck,
> 
> Guillaume
> 
> 
>> On 21 Jun 2022, at 19:38, Firdous Tarique > > wrote:
>> 
>> Hi
>> 
>> Sorry for the off topic question but can anybody please suggest how to 
>> dissolve Aluminium Fluoride in water ? I know it has little solubility in 
>> water but have seen people using AF3 for their structural studies (ADP + AF3 
>> as ATP analogue). Is there any other solvent where they have made AF3 
>> solution or is there some trick to dissolve it in water?.
>> 
>> Thanks
>> 
>> Firdous
>> 
>> To unsubscribe from the CCP4BB list, click the following link:
>> https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB=1 
>> 




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Re: [ccp4bb] Solubility of Aluminum Fluoride

[Firdous Tarique]

Thanks for your input. To avoid precipitation what about doing something
like this? For example, as we know: AlCl3 + 3NaF = AlF3 + 3NaCl

Therefore, adding 100mM AlCl3 plus 300mM NaF to get 100mM of AlF3 solution,
then directly using this 100 mM stock of AlF3 for the final dilution, say 1
or 2 mM in the reaction mix ? Or it is recommended to use AlCl3 and NaF
hoping to form AlF3 in the reaction mix.

Best

Faisal


On Tue, Jun 21, 2022 at 2:08 PM Guillaume Gaullier <
guillaume.gaull...@univ-tlse3.fr> wrote:

> Hi Firdous,
>
> I have read a fair amount of papers about ATP-dependent chromatin
> remodelers in which ADP-BeF3 is often cited. It is made as follows (quoting
> from one of these papers): "ADP-BeF3 was prepared as a 10x stock (10 mM
> ADP, 30 mM BeSO4, 150 mM NaF, 10 mM MgCl2), freshly before use."
>
> So the trick is to get all you need from a mixture of soluble compounds,
> instead of from only one poorly soluble salt. Not sure this will work the
> same with Al instead of Be, but it’s easy enough to try: use any
> known-soluble salt of Al instead of this BeSO4, and see if it works. NaF is
> extremely toxic, so use adequate protective measures (PPE, hood, etc.).
>
> Good luck,
>
> Guillaume
>
>
> On 21 Jun 2022, at 19:38, Firdous Tarique 
> wrote:
>
> Hi
>
> Sorry for the off topic question but can anybody please suggest how to
> dissolve Aluminium Fluoride in water ? I know it has little solubility in
> water but have seen people using AF3 for their structural studies (ADP +
> AF3 as ATP analogue). Is there any other solvent where they have made AF3
> solution or is there some trick to dissolve it in water?.
>
> Thanks
>
> Firdous
>
> --
>
> To unsubscribe from the CCP4BB list, click the following link:
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>
>
>



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Re: [ccp4bb] Solubility of Aluminum Fluoride

[Guillaume Gaullier]

Hi Firdous,

I have read a fair amount of papers about ATP-dependent chromatin remodelers in 
which ADP-BeF3 is often cited. It is made as follows (quoting from one of these 
papers): "ADP-BeF3 was prepared as a 10x stock (10 mM ADP, 30 mM BeSO4, 150 mM 
NaF, 10 mM MgCl2), freshly before use."

So the trick is to get all you need from a mixture of soluble compounds, 
instead of from only one poorly soluble salt. Not sure this will work the same 
with Al instead of Be, but it’s easy enough to try: use any known-soluble salt 
of Al instead of this BeSO4, and see if it works. NaF is extremely toxic, so 
use adequate protective measures (PPE, hood, etc.).

Good luck,

Guillaume


> On 21 Jun 2022, at 19:38, Firdous Tarique  wrote:
> 
> Hi
> 
> Sorry for the off topic question but can anybody please suggest how to 
> dissolve Aluminium Fluoride in water ? I know it has little solubility in 
> water but have seen people using AF3 for their structural studies (ADP + AF3 
> as ATP analogue). Is there any other solvent where they have made AF3 
> solution or is there some trick to dissolve it in water?.
> 
> Thanks
> 
> Firdous
> 
> To unsubscribe from the CCP4BB list, click the following link:
> https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB=1 
> 



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Re: [ccp4bb] Solubility of Aluminum Fluoride

[Michael Enos]

Hi Firdous,

When I have used ADP/AF3 as a transition state analogue in crystallography, I 
have always formed the ADP/AF3 complex in place by adding ADP, aluminum 
nitrate, and fluoride at a 1:1:6 ratio to my protein. I then used that mixture 
directly for crystallogenesis.

Best,
Michael

From: CCP4 bulletin board  On Behalf Of Firdous Tarique
Sent: Tuesday, June 21, 2022 12:38 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Solubility of Aluminum Fluoride

EXTERNAL MAIL
Hi

Sorry for the off topic question but can anybody please suggest how to dissolve 
Aluminium Fluoride in water ? I know it has little solubility in water but have 
seen people using AF3 for their structural studies (ADP + AF3 as ATP analogue). 
Is there any other solvent where they have made AF3 solution or is there some 
trick to dissolve it in water?.

Thanks

Firdous



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