By Harry Towbin (auth.), Biji T. Kurien, R. Hal Scofield (eds.)
Over the prior thirty years, the improvement of the Western blot has revolutionized the fields of biomedical learn and scientific diagnostics. In Protein Blotting and Detection: tools and Protocols, specialist researchers current various ideas according to the Western blot, offering special, without problems reproducible equipment, information, and possible choices at once and simply transferable to the laboratory surroundings. Chapters supply loads of adaptations at the subject matter of protein move to good aid through detection, featuring variations of conventional suggestions in addition to thoroughly unique equipment of protein blotting. Composed within the hugely profitable Methods in Molecular Biology sequence structure, each one bankruptcy incorporates a short advent, a listing of valuable fabrics, step by step tools, and a Notes part which stocks pointers on troubleshooting and keeping off identified pitfalls.
Innovative and hugely sensible, Protein Blotting and Detection: equipment and Protocols is a vital, hands-on consultant for all investigators who desire to carry those state of the art methods domestic to their laboratories.
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Additional resources for Protein Blotting and Detection: Methods and Protocols
Sample text
4. Nitrocellulose membrane (Gelman Sciences/Fisher Scientific, Dallas, TX, USA). 5. Polyvinylidene difluoride (PVDF) membrane (Millipore, Bedford, MA, USA). 6. , Farmingdale, New York, USA). 7. 7 mL (PGC Scientific, Frederick, MD, USA). 8. Super-Mixer (Curtin Matheson Scientific, USA). 3. Methods All procedures were performed at room temperature (unless indicated otherwise). 1. Cut a piece each of nitrocellulose and PVDF membranes and place them on top of a Whatman #3 filter paper cut slightly larger than the size of the membranes.
The IPG strip can be immediately subjected to second dimension of SDS-PAGE or be frozen at −20°C for later use. 6. The transfer time for diffusion blotting should be adjusted for gels with different thickness. 75 mm for 1 h and gel of 1 mm for 2 h. 7. After three washes with deionized water, the membrane can be air-dried and stored for later use. ) from the National Science Council, Republic of China. Simultaneous Immunoblotting Analysis with Activity Gel Electrophoresis 33 References 1. P. and Malamud, D.
Hiroyuki Matsumoto and Tryptic Peptide Purification Using Polyvinylidene Difluoride Membrane 43 Sadamu Kurono (Department of Biochemistry and Molecular Biology, Oklahoma City, OK, USA) for their help with mass spectrometry and analysis. References 1. , et al. (2006) Solvent-dependent metabolite distribution, clustering, and protein extraction for serum profiling with mass spectrometry. Anal Chem. 78, 743–752. 2. Fenselau, C. (1997) MALDI MS and strategies for protein analysis. Anal Chem. 69, 661A–665A.