Summary: Western Blotting, Western blot or immunoblotting (Western blotting was dubbed because it is similar to Southern blotting which was invented by and named after the inventor E. M. Southern) allows investigators to determine, with a specific primary antibody, the relative amounts of the protein present in different samples. Briefly, 1) Samples are prepared from tissues or cells that are homogenized in a buffer that protects the protein of interest from degradation; 2) The sample is separated using SDS-PAGE and then transferred to a membrane for detection; 3) The membrane is incubated with a generic protein (such as milk proteins) to bind to any remaining sticky places on the membrane. A primary antibody is then added to the solution which is able to bind to its specific protein; 4) A secondary antibody-enzyme conjugate, which recognizes the primary antibody is added to find locations where the primary antibody bound.
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Protocols
- Western Blotting Protocol (EnCor Biotechnology)
A basic nitrocellulose/PVDF protocol.
https://www.encorbio.com/protocols/blotting.htm
Added: Mon Feb 23 2009, Hits: 30910, Reviews: 0 Write review Cached - Western Blotting with Biotinylated Antibodies (BD Biosciences)
procedures including sample preparation from cell and tissue, quantification, electrophoresis, transfer, semi-dry transfer, blocking and troubleshootings.
https://www.bdbiosciences.com/support/resources/pro...
Added: Wed Aug 28 2002, Hits: 5245, Reviews: 0 Write review Cached - Western Blotting with Monoclonal Antibodies (BD Biosciences)
This is a detailed western protocol using monoclonal antibody for general use.
https://www.bdbiosciences.com/support/resources/pro...
Added: Tue May 14 2002, Hits: 8617, Reviews: 0 Write review Cached - Western Immunoblotting of Proteins (Millipore)
Western immunoblotting protocol for general use.
https://www.millipore.com/immunodetection/id3/weste...
Added: Tue Feb 17 2004, Hits: 11905, Reviews: 0 Write review Cached -
Western Blot
(R&D System)
Provides very nice and general protocol including Western Blot Procedure, immunostaining Procedure (standard and rapid), helpful hints and trouble shooting guide
https://www.rndsystems.com/western_blot_detail_obje...
Added: Tue May 14 2002, Hits: 15292, Reviews: 0 Write review Cached -
Western Blot
(Bjorkman Group, Howard Hughes Medical Institute at California Institute of Technology)
Describes sample preparation, SDS-PAGE, staining, transfer, blotting
https://www.its.caltech.edu/%7Ebjorker/Western_blot...
Added: Wed Aug 14 2002, Hits: 26848, Reviews: 0 Write review -
Western Blotting
(Howell Lab)
Procedures for preparation of cell lysate, gel, electrophoresis, transfer, antibody detection and stripping membranes.
https://cancer.ucsd.edu/howelllab/Western.html
Added: Tue May 14 2002, Hits: 13544, Reviews: 0 Write review Cached -
Anti-Phosphotyrosine Western Blotting
(Bowtell Lab Manual, Peter MacCallum Cancer Institute)
https://www.pmci.unimelb.edu.au/core_facilities/man...
Added: Tue May 14 2002, Hits: 540, Reviews: 0 Write review Cached -
MAP Kinase Gel Shift Assay
(Bowtell Lab Manual, Peter MacCallum Cancer Institute)
https://www.pmci.unimelb.edu.au/core_facilities/man...
Added: Tue May 14 2002, Hits: 302, Reviews: 0 Write review Cached -
Preparation of Brain Membrane Fractions for Western Blot Analysis
(BD PharMingen)
This is a standard protocol adapted from Rogers et al., 1991.
https://www.bdbiosciences.com/pharmingen/protocols/...
Added: Tue May 14 2002, Hits: 3818, Reviews: 0 Write review Cached -
Western Blot and Membrane Stripping
(Cowin Lab, New York University Medical Center)
Describes procedures for Western blotting and stripping blot for reblotting.
https://celljunctions.med.nyu.edu/lab/notprivate/me...
Added: Mon Mar 16 2009, Hits: 13890, Reviews: 0 Write review -
Western blot troubleshooting - Blank Film
(J. Rusty Bishop, PhD)
3 hr and 33 minute protocol to troubleshooting the major causes of blank film in a Western blot.
https://westernblot.org/western-blot-troubleshootin...
Added: Wed Aug 17 2011, Hits: 2652, Reviews: 0 Write review -
Western Blot with Platelet Protein
(Viral Immunity & Pathogenesis Group, the Université Catholique de Louvain)
Detailed western protocol devoted to detecting paltelet protein but can also be used for general purpose.
https://www.icp.ucl.ac.be/mexp/vipg_plwb.html
Added: Mon Mar 22 2004, Hits: 3275, Reviews: 0 Write review Cached -
Western Blots
(Dr. William H. Heidcamp, Biology Department, Gustavus Adolphus College)
https://homepages.gac.edu/~cellab/chpts/chpt4/ex4-8...
Added: Tue May 14 2002, Hits: 5285, Reviews: 0 Write review Cached -
Western Blotting
(Corbett Lab)
https://www.biochem.emory.edu/labs/acorbe2/western....
Added: Tue May 14 2002, Hits: 5485, Reviews: 0 Write review -
Western Blotting for Plant Tissue
(Preuss Lab, University of Chicago)
I. Prepare plant tissue samples: II. Pour Acrylamide Gel: III. Gel Electrophoresis: IV. Gel to Filter Blotting: V. Probing the Filter:
https://preuss.bsd.uchicago.edu/protocols/western.h...
Added: Mon Mar 16 2009, Hits: 1665, Reviews: 0 Write review -
Western Blotting Using Chemiluminscence
(Peter J. Hansen Lab)
https://www.animal.ufl.edu/hansen/protocols/blottin...
Added: Tue May 14 2002, Hits: 3044, Reviews: 0 Write review Cached -
Western Blotting with Alkaline Phosphatase Conjugates
(BD PharMingen)
Standard Western protocol describing protein preparation, gel preparation, blotting, blocking...
https://www.bdbiosciences.com/pharmingen/protocols/...
Added: Wed Aug 28 2002, Hits: 2209, Reviews: 0 Write review Cached -
Western Blotting with Horseradish Peroxidase Conjugates
(BD Biosciences)
Standard protocol in great detail.
https://www.bdbiosciences.com/support/resources/pro...
Added: Wed Aug 28 2002, Hits: 3006, Reviews: 0 Write review Cached -
Western Blotting with Rabbit Polyclonal Antibodies
(BD Biosciences)
Protocol for western blotting with polyclonal antibodies including troubleshootings.
https://www.bdbiosciences.com/support/resources/pro...
Added: Sun Aug 26 2012, Hits: 946, Reviews: 0 Write review Cached -
Western Procedures
(Gottschling Lab, Fred Hutchinson Cancer Research Center)
Protocols for running protein gel, transfer, staining, stripping and more.
https://labs.fhcrc.org/gottschling/General%20Protoc...
Added: Fri Jul 26 2002, Hits: 4595, Reviews: 0 Write review -
Yeast Lysates for Westerns
(Bowtell Lab Manual, Peter MacCallum Cancer Institute)
https://www.pmci.unimelb.edu.au/core_facilities/man...
Added: Tue May 14 2002, Hits: 182, Reviews: 0 Write review Cached
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