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Blotting

Blotting

(Science: molecular biology, procedure) general term for the transfer of protein, rna or dna molecules from a relatively thick acrylamide or agarose gel or to a paper like membrane (usually nylon or nitrocellulose) by capilliarity or an electric field, preserving the spatial arrangment. Once on the membrane, the molecules are immobilised, typically by baking or by ultra violet irradiation and can then be detected at high sensitivity by hybridisation (in the case of dna & rna) or antibody labelling (in the case of protein). Rna blots are called northern blots, dna blots, southern, protein blots, western.


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western blotting of DNA-binding protein of mammalian cell

... i have added other sample they ran properly but not this cell lysate... what could be problem..how to get a clear SDS-PAGE and western blotting....

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by drelamuruganvet
Mon Jan 19, 2015 9:39 am
 
Forum: Molecular Biology
Topic: western blotting of DNA-binding protein of mammalian cell
Replies: 0
Views: 394

use of a bacterial protease inhibitor cocktail with mammalia

Dear fellow biochemists, I need some advice on Western Blotting, more specifically the use of certain protease inhibitors with the RIPA cell lysis buffer and protease inhibitor cocktail. A Millipore technical representative recommended my using "539134 ...

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by rpwdoc
Tue Aug 26, 2014 10:08 pm
 
Forum: Cell Biology
Topic: use of a bacterial protease inhibitor cocktail with mammalia
Replies: 1
Views: 1928

Weird - shaped band on western blotting

Hi everyone, This is my first post in this forum. My name is Laila and currently working with the hepatoma cells AH109A to see the cell cycle regulation protein. Recently I got a weird-shaped band on my western blot. It is like an upside-down of smiling face in each lane. I use the same loading buff...

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by namakulaila
Thu Oct 17, 2013 6:17 am
 
Forum: Molecular Biology
Topic: Weird - shaped band on western blotting
Replies: 2
Views: 2449

protein fractionation

... at least of similar protein. If you have any, you just have to try different approaches and detect both your protein (enzymatic activity?, blotting...) and total protein and determine purification fold. Great for you would be the first chapters from methods in Enzymology 463 http://www.sciencedirect.com/science/bookseries/00766879/463

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by JackBean
Thu May 09, 2013 2:03 pm
 
Forum: Microbiology
Topic: protein fractionation
Replies: 4
Views: 3659

How can a protein without a G-site bind to GSH?

... the MALDI-TOF. In any case, because you will probably not have the protein in any database, you will have to create some gDNA/cDNA library and use blotting with (degenerated) probe.

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by JackBean
Sun Nov 25, 2012 1:08 pm
 
Forum: Molecular Biology
Topic: How can a protein without a G-site bind to GSH?
Replies: 11
Views: 4909
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