# Electroblotting

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Process in biochemistry

Schematic representation of a transfer stack.[1]

**Electroblotting** is a method in [molecular biology](/source/Molecular_biology)/[biochemistry](/source/Biochemistry)/[immunogenetics](/source/Immunogenetics) to transfer [proteins](/source/Proteins) or [nucleic acids](/source/Nucleic_acids) onto a membrane by using [PVDF](/source/PVDF) or [nitrocellulose](/source/Nitrocellulose), after [gel electrophoresis](/source/Gel_electrophoresis).[2][3] The protein or nucleic acid can then be further analyzed using probes such as specific [antibodies](/source/Antibodies), [ligands](/source/Ligand) like [lectins](/source/Lectin), or stains. This method can be used with all polyacrylamide and agarose gels. An alternative technique for transferring proteins from a gel is capillary blotting.

## Development

This technique was patented in 1989 by William J. Littlehales under the title "Electroblotting technique for transferring specimens from a polyacrylamide electrophoresis or like gel onto a membrane.[1][4]

## Electroblotting procedure

This technique relies upon current and a transfer buffer solution to drive proteins or nucleic acids onto a membrane. Following electrophoresis, a standard tank or semi-dry blotting transfer system is set up. A stack is put together in the following order from cathode to anode: sponge | three sheets of filter paper soaked in transfer buffer | gel | PVDF or nitrocellulose membrane | three sheets of filter paper soaked in transfer buffer | sponge. It is a necessity that the membrane is located between the gel and the positively charged anode, as the current and sample will be moving in that direction. Once the stack is prepared, it is placed in the transfer system, and a current of suitable magnitude is applied for a suitable period of time according to the materials being used.

Typically the electrophoresis gel is stained with [Coomassie brilliant blue](/source/Coomassie_brilliant_blue) following the transfer to ensure that a sufficient quantity of material has been transferred. Because the proteins may retain or regain part of their structure during blotting they may react with specific [antibodies](/source/Antibodies) giving rise to the term [immunoblotting](/source/Immunoblotting). Alternatively the proteins may react with [ligands](/source/Ligand) like [lectins](/source/Lectin) giving rise to the term [affinity](/source/Affinity_(pharmacology)) blotting.

## See also

- [Western blotting](/source/Western_blotting)

- [SDS-page](/source/SDS-page)

## References

1. ^ [***a***](#cite_ref-Dubey2014_1-0) [***b***](#cite_ref-Dubey2014_1-1) R C Dubey (2014). [*Advanced Biotechnology*](https://books.google.com/books?id=SKgrDAAAQBAJ&pg=PA199). S. Chand Publishing. pp. 199–. [ISBN](/source/ISBN_(identifier)) [978-81-219-4290-4](https://en.wikipedia.org/wiki/Special:BookSources/978-81-219-4290-4).

1. **[^](#cite_ref-Sheehan2013_2-0)** David Sheehan (30 April 2013). [*Physical Biochemistry: Principles and Applications*](https://books.google.com/books?id=AxxbYlsuZrwC&pg=PT206). John Wiley & Sons. pp. 206–. [ISBN](/source/ISBN_(identifier)) [978-1-118-68748-2](https://en.wikipedia.org/wiki/Special:BookSources/978-1-118-68748-2).

1. **[^](#cite_ref-Bansal2013_3-0)** M. P. Bansal (1 January 2013). [*Molecular Biology and Biotechnology: basic experimental protocols*](https://books.google.com/books?id=4rtpDzCi8NAC&pg=PA141). The Energy and Resources Institute (TERI). pp. 141–. [ISBN](/source/ISBN_(identifier)) [978-81-7993-379-4](https://en.wikipedia.org/wiki/Special:BookSources/978-81-7993-379-4).

1. **[^](#cite_ref-4)** ["United States Patent 4840714"](https://web.archive.org/web/20070929082908/http://www.patentstorm.us/patents/4840714.html). Archived from [the original](http://www.patentstorm.us/patents/4840714.html) on 2007-09-29. Retrieved 2007-05-24.

### External links

- Protein (Western) Blotting - Introduction to Antibodies [Membrane Transfer](https://web.archive.org/web/20070605124005/http://www.chemicon.com/Resource/ANT101/a2B.asp#Membrane)

- Detailed electroblotting to PVDF procedure - [Protein Chemistry Laboratory](https://web.archive.org/web/20070907163606/http://pcl.calabreso.com/protocols/pvdf_pro.html)

v t e Proteins: key methods of study Experimental Protein purification Green fluorescent protein Western blot Protein immunostaining Protein sequencing Gel electrophoresis/Protein electrophoresis Protein immunoprecipitation Peptide mass fingerprinting/Protein mass spectrometry Dual-polarization interferometry Microscale thermophoresis Chromatin immunoprecipitation Surface plasmon resonance Isothermal titration calorimetry X-ray crystallography Protein NMR Cryo-electron microscopy Freeze-fracture electron microscopy Bioinformatics Protein structure prediction Protein function prediction Protein–protein docking Protein structural alignment Protein ontology Protein–protein interaction prediction Assay Enzyme assay Protein assay Secretion assay Display techniques Bacterial display mRNA display Phage display Ribosome display Yeast display Super-resolution microscopy Photoactivated localization microscopy Vertico SMI

v t e Electrophoresis History of electrophoresis Techniques Affinity electrophoresis Agarose gel electrophoresis Capillary electrochromatography Capillary electrophoresis Dielectrophoresis Difference gel electrophoresis Discontinuous electrophoresis Electroblotting Electrochromatography Electrophoretic mobility shift assay Gel electrophoresis Immunoelectrophoresis Iontophoresis Isotachophoresis Moving-boundary electrophoresis Polyacrylamide gel electrophoresis Pulsed-field gel electrophoresis Temperature gradient gel electrophoresis Two-dimensional gel electrophoresis Applications DNA laddering DNA separation by silica adsorption Gel electrophoresis of nucleic acids Gel electrophoresis of proteins Serum protein electrophoresis Theory Electrical mobility Isoelectric focusing Journals Electrophoresis (journal) Category Commons Analytical Chemistry

v t e Molecular probes General Southern blot (DNA) Northern blot (RNA) Western blot (protein) Eastern blot (post translational modification) Interactions Southwestern blot (protein:DNA) Electrophoretic mobility shift assay (DNA:protein) Far-western blot (protein:protein) Far-eastern blot (lipid:post translational modification) Northwestern blot (RNA:protein)

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