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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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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: 973

Preparing cells for western blotting??

Dear All, i'm new in the cell culturing field and my Dr. asked me to prepare a western blotting protocol. its easy to understand each step, but im finding a diffeculty understanding the procedure in preparing cells before the protocol. what to do with the media n how ...

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by Halaqataruniversity
Fri Sep 07, 2012 2:53 pm
 
Forum: Cell Biology
Topic: Preparing cells for western blotting??
Replies: 1
Views: 1351

Radio-phosphate binding assay

... sonicate and resuspend in 10ml 10mM Tris-Cl buffer ph 7.0. 2) Pass 500ul protein through a native PAGE (I have combs this big). 3) Western blotting: transfer to an activated nitrocellulose/PVDF membrane). 4) Incubate blot in phosphate solution for 1 hour. 5) Develop blot in phospho-imaging ...

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by vader1941
Tue Jul 10, 2012 3:32 pm
 
Forum: Molecular Biology
Topic: Radio-phosphate binding assay
Replies: 0
Views: 909

fusion protein not expressing

... the GFP? What about trying the other end? Do you have any idea about structure of your protein? Did you try to detect your protein or GFP with blotting?

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by JackBean
Thu Feb 16, 2012 5:18 pm
 
Forum: Molecular Biology
Topic: fusion protein not expressing
Replies: 1
Views: 502

Southern Blott

It seems to me that you are talking about a regular gel before the blotting (I see no mention of probes or anything like that). In any case you want to measure the distance of migration from the bottom of the well to the middle of the band in the middle of ...

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by canalon
Wed Jan 04, 2012 2:10 pm
 
Forum: Genetics
Topic: Southern Blott
Replies: 4
Views: 1105
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