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POST ELECTROPHORETIC ANALYSIS Articles

Smeared Bands

By National Diagnostics | August 28, 2013 | Comments Off on Smeared Bands

Particulate in the sample: Standard loading buffers are often unable to completely dissolve complex samples-  fragments of bacterial cell walls, for example, can be carried over into the well.  Particulate material in the well blocks…

Blotches on Gel

By National Diagnostics | August 27, 2013 | Comments Off on Blotches on Gel

Blotches can be caused by overhandling of the gel, or by handling the gel without gloves. This can create uneven surfaces or protein deposits that can bind Coomassie to the gel. To minimize this problem,…

Uneven Staining

By National Diagnostics | August 27, 2013 | Comments Off on Uneven Staining

Uneven staining is almost always the result of insufficient agitation during staining.  The gel is initially more dense than the staining solution and will tend to sink to the bottom of the dish.  Some portions…

Faint Bands, High Background

By National Diagnostics | August 23, 2013 | Comments Off on Faint Bands, High Background

A high background which obscures the bands, or in combination with fainter than usual bands, indicates dye binding to the gel matrix, or contamination of the matrix with a dye-binding material (most often a protein).…

Faint bands, low background

By National Diagnostics | August 16, 2013 | Comments Off on Faint bands, low background

SDS in the staining solution:  Most users save and re-use their staining solutions.  This is fine for a couple of cycles, but over time enough SDS elutes from the gels to accumulate to significant levels…

Coomassie Blue Stain- Troubleshooting

By National Diagnostics | August 12, 2013 | Comments Off on Coomassie Blue Stain- Troubleshooting

If your gel doesn’t look like this one, click on the problem below to find the solution: Faint bands on a low background Learn More Faint bands on a high background Learn More Uneven Staining…

Mechanism of Immunostaining

By National Diagnostics | November 22, 2011 | Comments Off on Mechanism of Immunostaining

The highly specific binding interaction between antibodies and their unique antigens has been exploited to create sensitive and specific detection systems for proteins. An antibody can be raised and/or purified “against” (i.e. binding to) a…

Method for Western Blotting

By National Diagnostics | September 23, 2011 | Comments Off on Method for Western Blotting

ELECTROPHORESIS Prepare and run an SDS PAGE gel. Select a gel percent which will give the best resolution for the size of antigen being analyzed (if known). If the size is not known, a 12%…

Overview of Western Blotting

By National Diagnostics | September 19, 2011 | Comments Off on Overview of Western Blotting

Proteins can also be detected immunologically following electrophoresis, a technique known as Western blotting. This method relies on the fact that most epitopes (sites recognized by antibodies, generally comprising several amino acids) are still recognizable…

Enzyme Linked Immunosorbent Assay (ELISA)

By National Diagnostics | September 19, 2011 | Comments Off on Enzyme Linked Immunosorbent Assay (ELISA)

One of the most straightforward applications of immunological detection is the ELISA or enzyme-linked immunosorbent assay. In the simplest system, the bound antigen is probed with antibodies that carry covalently attached enzyme molecules. Antibody binding…