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10X PBS Recipe

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R Bishop
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Topic Started by R Bishop
on 11/14/2006 19:46 PM   
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How to make 10x Phosphate Buffered Saline, pH 7.4
(0.2 M phosphate, 1.5 M NaCl)

Materials
2.28 g NaH2PO4 (mw=120; 0.038M) or 2.62g NaH2PO4(H2O) (mw=137.99; 0.038M)
(Note: monobasic)
11.5g Na2HPO4 (mw=141.96; 0.162M)
(Note: dibasic)
43.84g NaCl

Make it up

Add NaH2PO4 and Na2HPO4 and NaCl to 450 ml dH2O.
Adjust pH to 7.4 by adding dilute (1N) HCl if necessary.
Bring the final volume to 500 ml with water.
Dilute 1:10 for final concentration.

Rb


"Everything should be made as simple as possible, but no simpler."
-- Einstein


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vilsy
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Posted By vilsy
on 3/15/2009 15:59 PM   
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Thanks a lot for this...

Vilay Khandelwal



Ivan
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Posted By Ivan
on 3/16/2009 10:58 AM   
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This may not be the place for this, but instead of making your own 10X PBS you can always buy it pre-made. This is why I bring this up: 


Reagent-wise, it will cost you about $6 to make 500 ml of 10X PBS (rough estimate). The same solution bought from vendors like Invitrogen would cost you between $20 and $30. The difference, about $10 to $20, is the amount of time you spend getting the reagents, weighing them, getting the distilled water, getting all of this into solution, getting the pH of the solution to the right pH, and very likely autoclaving the solution.


I am all for saving money, but at the same time I think it is important to consider every option. How much money is your time worth? Unfortunately a post-doc's time tends to be significantly undervalued. Plus you could argue that a bottle of 10X PBS is likely of higher quality than one made in the lab (nothing against a scientist's prowess in making solution, but from experience I can tell you that the autoclave is not always working as it should, the water filter has likely not been maintained for some time, and the balance well, it gets used a lot). 


My two cents :)

Ivan Delgado Orlic
Carlsbad, CA



Jen_Floyd
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Posted By Jen_Floyd
on 3/16/2009 15:50 PM   
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 I tend to agree with Ivan  here.  Since I moved to a company and all of the solutions and medias are stocked downstairs, my productivity has skyrocketed.  I easily get 10 times more done when I don' t have to worry about making reagents, troubleshooting what went wrong with reagents, etc.  The argument doesn't always work with every PI but it's a discussion worth having.



vilsy
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Posted By vilsy
on 3/16/2009 19:03 PM   
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Thanks for the suggestion. My prof is really old school and I am sure wont be interested in buying something when one can make it. We even write our own softwares and design our instruments and get it made in machine shops even when u can buy something similar at a competitive rate. I think the ideology is to make sure that her students know how to handle each and every aspect of their research and are not dependent on industry.


I will have a discussion though. Thanks anyways.

Vilay Khandelwal



TheFFM
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Posted By TheFFM
on 4/1/2009 19:58 PM   
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vilsy said:


Thanks for the suggestion. My prof is really old school and I am sure wont be interested in buying something when one can make it. We even write our own softwares and design our instruments and get it made in machine shops even when u can buy something similar at a competitive rate. I think the ideology is to make sure that her students know how to handle each and every aspect of their research and are not dependent on industry.


I will have a discussion though. Thanks anyways.



 


there is a distinct difference in the level of technical acumen needs to acquire to make up a liter of PBS and to assemble a patch clamp amplifier!  Good luck with your persuasion!

"Nothing shocks me.....I'm a scientist!"
Indiana Jones


Last edited Apr 01, 2009, 17:58 PM by TheFFM

chrisg
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Posted By chrisg
on 11/11/2009 5:01 AM   
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This probably sounds like a stupid question but if I wanted to make a 10x PBS solution with 0.1M phosphate do I simply half the weight of the phosphate components given?



BSharp78
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Posted By BSharp78
on 11/24/2009 7:34 AM   
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Hi all,

I noticed that there is no KCl in this recipe.  Is it a necessary ingredient?  PS- I agree with buying the reagents, but I would like to know how to make it anyways.  Thanks.



Neznika
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Posted By Neznika
on 12/8/2009 7:42 AM   
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Sure. KCl is not a necessary ingredient in this buffer. And KH2PO4 is not a necessary here. Because there is the recipe of 0.2 M phosphate, 1.5 M NaCl PBSaline. I guess you are talking about PBSaline recipe kind of:

o        137 mM NaCl /     2.7  mM KCl /    10  mM Na2HPO4 / 2  mM KH2PO4/ pH 7.4
PBS is a common abbriviation of Phosphate Buffered Saline and Phosphate Buffer Solution (buffers  w/o salts ). Some people use different lable for NaH2PO4/Na2HPO4 buffers - SPB (Sodium Phosphate Buffer).

Exemples of others PBSalines you may look here:
herehttp://www.ihcworld.com/_protocols/washing_buffers/pbs.htm

 


Last edited Dec 08, 2009, 18:02 PM by Neznika

Neznika
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Posted By Neznika
on 12/8/2009 7:57 AM   
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It's Ok, Chris. At 25degC for pH 7.4 and 100ml the recipe is:
 40.5 ml of 0.2M Na phoshate dibasic + 9.5ml of 0.2M Na phos. monobasic. Adjust to 100ml.

Look details for others pH in "Data of Biochemical Reaserch" of Rex M. Dawson and others... (I have it as Third Edition, Clarendon Press, Oxford, 1986).



cambio
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Posted By cambio
on 12/17/2009 15:59 PM   
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Be careful when you store this. I find that storing 10x PBS at 4 degrees C and you will get precipitation at the bottom. I also filter my 10x stock.



Neznika
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Posted By Neznika
on 12/22/2009 14:55 PM   
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to cambio.
10xPBS couldn't be stored at 4degC. I guess it couldn't be stored at 10degC or 15 too because the precipitation of solts. So just autoclave 10xPBS and keep it benchtop. Plus (or minus) - don't filter the precipitate as you will decrease the concentration of solts in the stock. If the precipitation was, keep the stock bench top at RT/ couple hours (or a little bit hit it or autoclave) and the precipitate will be dissolved.
good luck


Last edited Dec 28, 2009, 8:38 AM by Neznika

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