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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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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: 838

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: 2073

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: 2566

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: 2107

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: 1295
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