Electroblotting

Electroblotting

Electroblotting is a method in molecular biology/biochemistry/immunogenetics to transfer proteins or nucleic acids onto a membrane (generally PVDF or nitrocellulose) after gel electrophoresis. The protein or nucleic acid can then be further analyzed using probes such as specific antibodies 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.

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." [ [http://www.patentstorm.us/patents/4840714.html United State Patent 4840714] ]

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 cathode, as the current and sample will be moving in that direction (the protein will have a negative charge due to interactions with SDS). 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 Blue following the transfer to ensure that a sufficient quantity of material has been transferred.

References

Footnotes

Notations

* Information on Electroblotting [http://www.serva.de/products/sheets/blotting.shtml Serva Electrophoresis]
* Protein (Western) Blotting - Introduction to Antibodies [http://www.chemicon.com/resource/ANT101/a2B.asp#Membrane Membrane Transfer]

ee also

* Western blotting
* SDS-page
* Gel electrophoresis

Related Links

* Detailed electroblotting to PVDF procedure - [http://pcl.calabreso.com/protocols/pvdf_pro.html Protein Chemistry Laboratory]


Wikimedia Foundation. 2010.

Игры ⚽ Нужен реферат?

Look at other dictionaries:

  • electroblotting — см. blotting. (Источник: «Англо русский толковый словарь генетических терминов». Арефьев В.А., Лисовенко Л.А., Москва: Изд во ВНИРО, 1995 г.) …   Молекулярная биология и генетика. Толковый словарь.

  • electroblotting — noun A form of electrophoresis in which the separated materials are transferred onto a membrane …   Wiktionary

  • Elsayed Elsayed Wagih — Infobox Scientist image width = 150px name = Elsayed Elsayed Wagih birth date = November 21st, 1946 birth place = Alexandria, Egypt flagicon|EGY residence = Alexandria, Egypt nationality = Egyptian field = Virology and Biotechnology work… …   Wikipedia

  • Gel electrophoresis — apparatus – An agarose gel is placed in this buffer filled box and electrical field is applied via the power supply to the rear. The negative terminal is at the far end (black wire), so DNA migrates toward the camera. Classification… …   Wikipedia

  • SDS-PAGE — PAGE redirects here. For other uses, see Page (disambiguation). Picture of an SDS PAGE. The molecular marker is in the left lane SDS PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis, describes a collection of related techniques… …   Wikipedia

  • Western blot — The western blot (alternatively, immunoblot) is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native or denatured proteins by the length of the… …   Wikipedia

  • Electroelution — is a method used to extract a nucleic acid or a protein sample from an electrophoresis gel by applying a negative current in the plane of the smallest dimension of the gel, drawing the macromolecule to the surface for extraction and subsequent… …   Wikipedia

  • RBBP4 — Retinoblastoma binding protein 4, also known as RBBP4, is a human gene. PBB Summary section title = summary text = This gene encodes a ubiquitously expressed nuclear protein which belongs to a highly conserved subfamily of WD repeat proteins. It… …   Wikipedia

  • HNRPA2B1 — Heterogeneous nuclear ribonucleoprotein A2/B1, also known as HNRPA2B1, is a human gene. PBB Summary section title = summary text = This gene belongs to the A/B subfamily of ubiquitously expressed heterogeneous nuclear ribonucleoproteins (hnRNPs) …   Wikipedia

  • P4HB — Prolyl 4 hydroxylase, beta polypeptide PDB rendering based on 1bjx …   Wikipedia

Share the article and excerpts

Direct link
Do a right-click on the link above
and select “Copy Link”