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Protein Science Post Questions and Discuss Protein Structure and Function, Protein Localization etc.. Please do not post Proteomic questions here, there is a Proteomic Forum below.


DTT Protein Extraction

DTT Protein Extraction - Protein Science

DTT Protein Extraction - Post Questions and Discuss Protein Structure and Function, Protein Localization etc.. Please do not post Proteomic questions here, there is a Proteomic Forum below.


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  #1  
Old 07-23-2007, 01:41 PM
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Smile DTT Protein Extraction



One question which I got from an email from a fellow researcher:


Hi,
which reagent could replace DTT in Protein extraction?????? In sample buffer
?

thanks
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Old 07-23-2007, 02:46 PM
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Default Re: DTT Protein Extraction

BME-beta mercaptoethanol
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Old 09-09-2007, 08:16 AM
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Default Re: DTT Protein Extraction

hi,

danfive is right. 2-marcaptoethanol is the replacement of DTT (Dithiothreitol / a Cleland's reagent . But i would suggest u to use DTT bcoz 2-marcaptoethanol is dangerous while DTT is not. Moreover, If you are working on a mixture of unknown proteins then some researchers prefer to use a combinition of DTT+marcaptoethanol.
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Old 10-09-2007, 04:47 PM
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Default Re: DTT Protein Extraction

Quote:
Originally Posted by admin View Post

Hi,
which reagent could replace DTT in Protein extraction?????? In sample buffer
?
Just learned that these reducing agents can be substituted for DTT (performance would be sample-dependent, as usual ) TBP-tributylphosphine, TCEP-tris (2-carboxyethyl)phosphine.
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Old 10-10-2007, 06:57 AM
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Default Re: DTT Protein Extraction

thank you very helpful for all.

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Old 10-25-2007, 07:12 AM
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Default Re: DTT Protein Extraction

Hi!
which difference between ampholytes and carrier ampholytes and his role this is my first question and the second is can I use the IPG-buffer with pH 3-10 for exemple and the strips with another pH?
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Old 10-29-2007, 04:00 PM
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Default Re: DTT Protein Extraction

Quote:
Originally Posted by walid View Post
Hi!
which difference between ampholytes and carrier ampholytes and his role this is my first question and the second is can I use the IPG-buffer with pH 3-10 for exemple and the strips with another pH?
Carrier ampholytes are just zwitterionic compounds that will separate into a 3-10 pH/pI gradient under isolectric separation. Yes you can use pH 3-10 ampholytes in the buffer with an IPG strip that can be 4-7, 7-10, anything that falls within the 3-10 pH range.
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Old 10-30-2007, 10:34 AM
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Default Re: DTT Protein Extraction

I have more questions:
1-which reagent could replace chaps in protein extraction?
2- the quantity of the sample (I want it in µl and in µg) that I used it with rehydratation of strip 7 cm or strip 24 cm overnight and I must diluted it or no and which dilution is better?
3-the composition of solubilsation buffer that I can use it with all type of fruit?
4- Which best staining solution with comassie blue or silver staining with total protein extraction i.e. I don't know the type of protein if it is acidic or basic protein?
5-when I worked in extraction of total protein which type of strip is better can used it i.e. with pH 3-10 NL or another type of strip?
6-concerneing the gel which better the dabble gels i.e. stacking gel + gel de concentration or one gel?
7-the colour of the gel is not clear the problem is in staining solution or what?
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Old 07-30-2008, 02:19 PM
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Default Re: DTT Protein Extraction

[QUOTE=admin;4401]One question which I got from an email from a fellow researcher:


which reagent could replace chaps in protein extraction?
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Old 12-08-2011, 10:26 AM
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Default Re: DTT Protein Extraction

I agree with danfive.
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