Download Immunocytochemical Methods and Protocols by Su-Yau Mao, Lorette C. Javois, Ute M. Kent (auth.), Lorette PDF

By Su-Yau Mao, Lorette C. Javois, Ute M. Kent (auth.), Lorette C. Javois (eds.)

Lorette Javois' well timed new second variation revises and updates her commonly acclaimed selection of step by step immunocytochemical tools, one who is now utilized in many organic and biomedical study courses. The tools are designed for researchers and clinicians who desire to visualize molecules in plant or animal embryos, tissue sections, cells, or organelles. as well as state-of-the-art protocols for purifying and getting ready antibodies, gentle microscopic research, confocal microscopy, FACS, and electron microscopy, this revised version includes many new equipment for making use of immunocytochemical strategies within the medical laboratory and together with in situ hybridization.

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2. Formaldehyde/Paraformaldehyde Formaldehyde is a colorless gas that is soluble in water (3). Commercial aqueous preparations of formalin contain 37–40% w/w solubilized gas. 05%) and 10–15% methanol, which is added to prevent the polymerization of formaldehyde into paraformaldehyde (3,11). Methanol and formic acid make these solutions an unacceptable fixative for fine structures (9). Paraformaldehyde is a polymerized form of formaldehyde that dissociates at 60°C and neutral pH. Freshly prepared solutions of paraformaldehyde are preferred for most immunochemical procedures because they provide a fixative free of extraneous additives and are usually the conservative fixatives of choice when beginning the development of a fixation protocol (3,5).

Bloodgroup substance A coupled to agarose as an immunosorbent. Biochim. Biophys. Acta 362, 567–574. 7. Affinity Chromatography–Principles and Methods. (1983) Pharmacia-LKB, Ljungfoeretagen, Oerebro AB, Sweden. 8. , and Ottosson, T. (1983) Desorption of immunoglobulins from Protein A-Sepharose Cl-4B under mild conditions. J. Immunol. Methods 64, 1–6. 9. Ey, P. , Prowse, S. , and Jemkin, C. R. (1978) Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using Protein A-Sepharose.

1–201. 10. Goldenthal, K. , Chen, J. , August, J. , and Willingham, M. C. (1985) Postfixation detergent treatment for immunofluorescence suppresses localization of some integral membrane proteins. J. Histochem. Cytochem. 33, 813–820. 11. Kiernan, J. A. (1981) Histological and Histochemical Methods. Theory and Practice. Pergamon, Oxford, UK. 12. Ito, S. and Karnovsky, M. J. (1968) Formaldehyde-glutaraldehyde fixatives containing trinitro compounds. J. Cell Biol. 39, 168a. 13. Newman, G. , and Williams, E.

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