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Cell Lysis Buffer


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6 replies to this topic

#1 bioJ

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Posted 29 October 2010 - 05:02 AM

Can someone give me a recipe for a cell lysis buffer...here is what I"m trying to do:

Detect Ras in HEK293 cells by western.

Thanks!

#2 madrius1

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Posted 29 October 2010 - 06:07 AM

1% Triton x-100
50 mM Tris pH7.4
150 mM NaCl
1 mM EDTA
Protease/phosphatase inhibitors

Should do the trick.

#3 bioJ

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Posted 29 October 2010 - 06:40 AM

great thanks! What is the difference between using tritonx or sds as the detergent?

#4 Halfro22

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Posted 29 October 2010 - 01:51 PM

I think it is their degree of "harshness", whatever that means. What method are you using for protein quantitation?

#5 mdfenko

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Posted 04 November 2010 - 08:05 AM

triton is non-ionic and sds is ionic
talent does what it can
genius does what it must
i do what i get paid to do

#6 castleinthesky

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Posted 01 March 2012 - 07:07 PM

1X Cell Lysis Buffer:
20 mM Tris-HCl (pH 7.5)
150 mM NaCl
1 mM Na2EDTA
1 mM EGTA
1% Triton
2.5 mM sodium pyrophosphate
1 mM b-glycerophosphate
1 mM Na3VO4
1 μg/ml leupeptin
some protease inhibitor added will make it better~

#7 bob1

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Posted 02 March 2012 - 12:56 PM

1X Cell Lysis Buffer:
20 mM Tris-HCl (pH 7.5)
150 mM NaCl
1 mM Na2EDTA
1 mM EGTA
1% Triton
2.5 mM sodium pyrophosphate
1 mM b-glycerophosphate
1 mM Na3VO4
1 μg/ml leupeptin
some protease inhibitor added will make it better~

Castleinthesky, it might be worth you checking when these posts were last posted in, this one is well over a year old...




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