By Nathan P. Kaplan, Nathan P. Colowick, John J. Langone, Helen Van Vunakis
The significantly acclaimed laboratory usual, Methods in Enzymology, is among the so much hugely revered guides within the box of biochemistry. on the grounds that 1955, each one quantity has been eagerly awaited, usually consulted, and praised by means of researchers and reviewers alike. The sequence includes a lot fabric nonetheless suitable this present day - really a vital booklet for researchers in all fields of existence sciences
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Extra resources for Immunochemical Techniques - Part C
The precipitate is immediately centrifuged at 16,000 g for 30 min at 20°. 001 M NAN3. 55 serum volume portion of ammonium sulfate. 5 mg/ml. As an alternative to antibody preparations, receptors from tissues that 2r A. J. Portmann, S. A. Levison,and W. B. Dandliker,lmmunochemistry 12, 461 (1975). M. B. Sigel,W. P. Vanderlaan,E. F. Vandedaan,and U. J. Lewis,Endocrinology 106, 92 (1980).  FLUORESCENCE POLARIZATION ASSAYS 27 are targets for hormones, drugs, toxins, viruses, and bacteria may sometimes prove advantageous in assays.
Labeling the antibody with quencher has the effect of reversing the enhancement phenomenon. The result is weak overall antibody-induced quenching superimposed on significant sample to sample variation caused by the serum effect. The problems associated with fluorescence enhancement can be addressed by appropriate modification of the fluorescer conjugate to avoid internal quenching. In the case of triiodothyronine, a conjugate with fluorescer 10b that showed fluorescence enhancement by serum of up to about 20% became quite well behaved when attached to an aminosubstituted dextran, possibly because of steric protection from nonspecific binding by serum proteins.
M. Brinkley, and R. F. Zuk, in "Immunoassays: Clinical Laboratory Techniques for the 1980s " ( R . M. Nakamura, W. R. Dito, and E. S. ). Liss, New York (1980). , Dayton, Ohio. < ~4 < ? z X X ? X ? X ,~~K ? ~ ~-~ ~ ~ ~z i. ) ,... 26 IMMUNOASSAYMETHODS  Antibody or Receptor Preparation The qualities desired in an antibody or receptor preparation to be used in an assay include (1) high binding affinity, (2) uniformity of binding affinity, (3) high specificity for the molecular structure being assay, (4) low adventitious fluorescence, (5) stability on storage, and (6) low cost.
Immunochemical Techniques - Part C by Nathan P. Kaplan, Nathan P. Colowick, John J. Langone, Helen Van Vunakis