MAP Kinase Signaling Protocols (Methods in Molecular Biology by Rony Seger

By Rony Seger

Show description

Read Online or Download MAP Kinase Signaling Protocols (Methods in Molecular Biology Vol 250) PDF

Best biology books

Translation Initiation: Extract Systems and Molecular Genetics

For over fifty years the tools in Enzymology sequence has been the seriously aclaimed laboratory commonplace and some of the most revered courses within the box of biochemistry. The hugely suitable fabric makes it a vital ebook for researchers in all fields of lifestyles and similar sciences.

Evolutionary biology of ostracoda: its fundamentals and applications: Proceedings of the Ninth International Symposium on Ostracoda

There are various aggressive works out there bearing on evolutionary biology, yet this quantity is kind of specified in its idiographic point targeting Ostracoda seen from a variety of disciplines, a while and environments. The ebook bargains with a variety of strains of idiographic biology and palaeontology of Ostracoda and nomothetic trials focusing strongly on evolutionary biology.

Liposomes: Methods and Protocols, Volume 2: Biological Membrane Models

With approximately 100 years of extensive learn, lipids have confirmed to be an essential and ever-more-promising zone of phone organic learn. In Liposomes: tools and Protocols, major specialists within the similar fields discover state of the art experimental tools regarding all features of lipids as crucial elements of the mobile membrane.

Fungi : biology and applications

Fungi: Biology and functions, moment Edition€provides a complete therapy of fungi, overlaying biochemistry, genetics and the scientific and fiscal importance of those organisms at introductory point. € without past wisdom of the topic assumed, the hole chapters supply a large evaluation of the fundamentals of fungal biology, particularly the body structure and genetics of fungi and in addition a brand new bankruptcy at the software of genomics to fungi.

Additional info for MAP Kinase Signaling Protocols (Methods in Molecular Biology Vol 250)

Sample text

10% Ammonium persulfate (APS) (#161-0700; Bio-Rad). 9. 3. 10. 005% bromophenol blue. 11. Destaining solution: 15% isopropanol, 7% acetic acid. 3. Western Blot Analysis 1. 2. 3. 4. 5. 6. 7. 8. 9. Transfer apparatus. 8. Nitrocellulose membrane (Protran BA 85; Schleicher & Schuell). Whatman paper (3 mm). 05% Tween-20. Blocking solution: 2% (w/v) BSA in washing buffer. , monoclonal antidiphospho-ERK [M-8159] and polyclonal anti–general ERK [M-5670] from Sigma, Israel), and secondary antibody (alkaline phosphatase [AP]–or horseradish peroxidase [HRP]–conjugated antimouse or antirabbit Fab antibodies from Jackson) diluted in washing buffer to appropriate dilutions.

End the phosphorylation reactions by adding 10 µL of 4X sample buffer to each tube. Boil, centrifuge (1 min at 14,000g), and load the supernatants on a 15% SDS-PAGE gel. 5 cm from the bottom of the gel, stop the current. To remove the excess free radiolabeled ATP, which migrates just in front of the bromophenol blue, cut out the part of the gel below the dye. This will considerably reduce the amount of radioactivity in the gel. 3.. Wash briefly with distilled water and let dry. An alternative way would be to stain, destain, and dry the gel on a Whatman 3-mm paper, but this procedure does not allow further detection of proteins in the gel as described for the immunoprecipitated ERKs in step 16.


Download PDF sample

Rated 4.55 of 5 – based on 37 votes