Immunochemical Protocols (Methods in Molecular Biology Vol by Margaret M. Manson

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By Margaret M. Manson

Highlights fresh advances in immunochemical thoughts and their impression on uncomplicated examine and scientific medication, combining the forces of specialists in a variety of subspecialties to hide a wealth of knowledge.

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9. 0 and dilute to 10 mMor 50 mMas required. 10. 9% NaCl. 11. Acetone. 12. Biogel PlO column. 13. Dithiothreitol (DTI’), 100 mA4. 14. 20 mL P30 (Bio-Rad) column. 3. 1. Glutaraldehyd-e Coqjugation Procedure (see Note 1) 1. Weigh out the peptide and an equal weight of KLH or thyroglobulin carrier. This gives an approximate ratio of 40-150 molecules of peptide to each molecule of carrier (2 mg of peptide per animal is ample). , for a 15 amino acid peptide, use 3 mg of peptide for each 10 mg of PPD).

Springer-Verlag, Berlin. Bodanszky, M. (1988) Ptprde Chemtshy. A FbcticalA&bvach. Springer-Verlag, Berlin. Atherton, E. and Sheppard, R. C. (1984) Sold Phase Pepa& Syvtthcw: A E4achual Ap jnnach. lRL Press, Oxford Lachmann, P. , Vyakamam, A. and Evan, G. I. (1984) Raismg annbodies by coupling peptides to PPD and immumsing BCGsensitised animals. CIBA Foundation Symposium. John Wiley, Chichester CHAPTER5 Production and Characterization of Antibodies Against Synthetic Peptides David C. Hancock and Gerard I.

59. 30 cm x 1 cm Column of Sephadex G-75 equilibrated with UB buffer. 60. 5-20 mL. 3. 1. , Chapter 4). 1. , pipet 250 PL of each into a 5-mL glass beaker on a magnetic stirrer. Small fleas can be made from pieces of paper clip sealed in polythene tubing by heating. Add 5 PL of 25% glutaraldehyde and continue stirring for 15 min at room temperature. Block excess glutaraldehyde by adding 100 PL of 1Mglycine and stirring for a further 15 min. Use directly or dialyze overnight against PBS and store at -2OOC.

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