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stacking gel


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4 replies to this topic

#1 SF_HK

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Posted 10 August 2009 - 01:48 AM

Hi,

I made a solution of 1.5M tris with a required pH of 8.8. Howver, when I was adjusting the pH, I added too much HCL initially initially and had to adjust it by adding some alkaline. Is that ok? Will it affect my western?

#2 Gerard

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Posted 10 August 2009 - 10:46 AM

No problem 1.5M has not to be very precise
Ockham's razor
Pluralitas non est ponenda sine necessitate
-- "You must assume no plural without necessity".

#3 cellcounter

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Posted 10 August 2009 - 11:48 AM

Hi,

I made a solution of 1.5M tris with a required pH of 8.8. Howver, when I was adjusting the pH, I added too much HCL initially initially and had to adjust it by adding some alkaline. Is that ok? Will it affect my western?

If I were you, I would throw away what you have prepared and make a brand new reagent. Be safe than sorry later.

M and pH of solutions do matter for obtaining good resolution. Evne if you don't run into problems with your current protein, your may, in the future, when you use the compromised reagent, forgetting how it was made.

#4 mdfenko

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Posted 11 August 2009 - 09:21 PM

also, the separation gel buffer is 1.5M, Ph 8.8. the stacking gel buffer is 1M, pH 6.8 (the pH being the more important parameter, volume used can be adjusted for differing concentration).
talent does what it can
genius does what it must
i do what i get paid to do

#5 T C

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Posted 14 August 2009 - 12:21 AM

Hey,

Just make sure that the pH is correct and the molarity doesn't change. It will be okay.

Best,
TC

Hi,

I made a solution of 1.5M tris with a required pH of 8.8. Howver, when I was adjusting the pH, I added too much HCL initially initially and had to adjust it by adding some alkaline. Is that ok? Will it affect my western?






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