By Harry Towbin (auth.), Biji T. Kurien, R. Hal Scofield (eds.)
Over the earlier thirty years, the improvement of the Western blot has revolutionized the fields of biomedical examine and clinical diagnostics. In Protein Blotting and Detection: equipment and Protocols, specialist researchers current a variety of concepts in accordance with the Western blot, offering certain, simply reproducible equipment, advice, and possible choices without delay and simply transferable to the laboratory environment. Chapters supply a lot of diversifications at the subject matter of protein move to good help by means of detection, providing variations of conventional recommendations in addition to thoroughly unique tools of protein blotting. Composed within the hugely profitable Methods in Molecular Biology sequence layout, every one bankruptcy features a short advent, a listing of beneficial fabrics, step by step tools, and a Notes part which stocks pointers on troubleshooting and averting recognized pitfalls.
Innovative and hugely useful, 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 strategies domestic to their laboratories.
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Additional info for Protein Blotting and Detection: Methods and Protocols
1% Triton X-100 and 7% acetic acid. 7. Staining for 2-DE (17) 1. 20% TCA solution. 2. 2 mL of glutardialdehyde (25%) to 40 mL of the stock solution prior to use (see Note 4). 3. 25% silver nitrate as stock solution and add 20 μL of formaldehyde (37%) to 40 mL of the stock solution prior to use (see Note 4). 4. 5% sodium carbonate as stock solution and add 10 μL of formaldehyde to 40 mL of the stock solution prior to use (see Note 4). 5. 4% Acetic acid. 8. Immunoblotting (18) 1. Phosphate buffered saline (PBS).
And De Loof, A. (1982) Vacuum-blotting: a new simple and efficient transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to NC. FEBS Lett 145, 369–372. Chapter 4 Simultaneous Immunoblotting Analysis with Activity Gel Electrophoresis and 2-D Gel Electrophoresis Der-Yen Lee and Geen-Dong Chang Summary Diffusion blotting method can couple immunoblotting analysis with another biochemical technique in a single polyacrylamide gel, however, with lower transfer efficiency as compared with the conventional electroblotting method.
The purified neutrophil membranes were solubilized and passed over an anti-Ro 60 affinity chromatography column. The proteins eluted from the column were analyzed on SDS PAGE and transferred nonelectrophoretically to nitrocellulose membrane (24). The membrane was immunoblotted with anti-Ro 60 sera. The residual proteins on the gel were stained with Coomassie blue (25), and the immunoblotted band was compared with the Coomassie blue-stained gel and identified. The target protein band was excised and subjected to in-gel tryptic digestion.
Protein Blotting and Detection: Methods and Protocols by Harry Towbin (auth.), Biji T. Kurien, R. Hal Scofield (eds.)