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refers to a number of similar techniques that facilitate the analysis of biological molecules. Initially, mixtures of proteins or dna or rna fragments are separated by electrophoresis through a polymeric gel (most commonly agarose or polyacrylamide). The separated components are then transferred to a membrane where they are immobilised and can be analysed and visualised by a variety of techniques, including antibody binding and dna hyrbridisation. Some of the various types of blots are briefly described below. Southern blot - The gel is a dna electrophoresis system and the molecules transferred are dna molecules. Northern blot - similar to the southern blot, but used to analyse rna. Western blot - The molecules analysed in a western blot are proteins, separated by gel electrophoresis, then identified by reacting them with an antibody. Southwestern blot - this is a variant of the southern blot used to find protein molecules that stick to dna molecules. Dot blot. Dna, rna, or protein are dotted directly onto the membrane support, so that they form discrete spots.

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Animal-Derived Laboratory Reagents

I have a question about animal-derived laboratory reagents and products. I’m working in a strictly in-vitro lab that focuses mainly on western blots and flow cytometry. I was wondering the cost to animal life associated with these reagents. These include: FBS (fetal bovine serum) & BSA (bovine ...

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by southtowns18
Thu Dec 06, 2012 6:13 pm
Forum: General Discussion
Topic: Animal-Derived Laboratory Reagents
Replies: 0
Views: 1589

Western Blot TroubleShooting

Dear all, I need some help with Western Blots. I am trying to detect a transcription factor that is expressed at pretty low levels endogenously in cell lines. Does anyone have any suggestions? -I use RIPA buffer with protease inhibitor to harvest ...

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by mutant11
Fri Nov 18, 2011 10:17 pm
Forum: Molecular Biology
Topic: Western Blot TroubleShooting
Replies: 7
Views: 4028

BN-PAGE of purified membrane associated protein

... %) because the protein seems to form high aggregated complexes. There is still one problem. I got protein smears so that I cannont perform Western blots. Attached you find an example. In the first two lines I loaded two different markers; first is BSA, second JackBean Urease. Then different protein ...

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by Papaver
Wed Nov 16, 2011 12:28 pm
Forum: Molecular Biology
Topic: BN-PAGE of purified membrane associated protein
Replies: 7
Views: 2821

Western Blotting of small molecular weight

Hi, I am working on a project in which I am doing Western Blots to look at the expression of a low molecular weight protein (15 kDa) in various brain regions. I have been using a 0.2 uM nitrocellulose membrane for my transfers and I have been transferring ...

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by Homie7
Thu Sep 08, 2011 11:46 am
Forum: Molecular Biology
Topic: Western Blotting of small molecular weight
Replies: 4
Views: 7554

probing mRNA and RT-PCR products

... right or a text or website where I could find the answer? Thanks Questions refering to the image 1) what size mRNA would be detected on northern blots hybridised with PROBE1 - (1kb? exon 1 only) 2) what size mRNA would be detected on northern blots hybridised with PROBE2 - (1kb? exon 2 only) ...

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by SarahTessler
Thu Jun 09, 2011 4:22 am
Forum: Molecular Biology
Topic: probing mRNA and RT-PCR products
Replies: 4
Views: 2873
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