By Shirley J. Gee
, Pages v-vii
, Page viii
, Page ix
Section 1 - Introduction
, Pages 1-6
Section 2 - getting ready the laboratory
, Pages 7-8
Section three - Laboratory considerations
, Pages 9-13
Section four - Immunoassay tutorials for chosen environmental analytes
, Pages 14-53
Section five - Tutorials for help techniques
, Pages 54-85
Section 6 - word list of usual phrases in Immunoassay
, Pages 86-90
Appendix I - functionality insurance for air displacement pipettes
, Pages 91-102
Appendix II - functionality log and function try out worksheets for air displacement pipettors
, Pages 103-106
Abbreviations and acronyms
, Page 107
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Extra resources for Environmental Immunochemical Analysis Detection of Pesticides and Other Chemicals. A User's Guide
Add 100 pL to each well of a high binding ELISA microtiter plate. Cover the microtiter plate with a plate sealer and incubate overnight at 4~ See Note 3. Wash the coated microtiter plate 5X with P BS-Tween/Azide and tap dry. The wash procedure involves flooding each well with I1~ Enzyme-labelled ]~ 1 Rabbit / ~ anti-rabbit antibody / H Competing free : n t t : b H ~ n H H Hapten~ _~ H protein H--~PROTEINT conjugate Well of polystyrene96-well plate Figure 9. Schematic of the anitgen-coated plate ELISA formal The fines represent wash steps.
I 11 ll I Add 50 I~L of standard or sample from the mixing plate to each well of the coated plate. Add 50 ILL of Antibody #2114 diluted 1/40,000 in PBS-Tween/Azide to each well of the coated plate, except those wells that serve as no-antibody blanks. ) In the wells that serve as blanks, replace the antibody with buffer. See Note 6. Cover the coated plate with a plate sealer and incubate 1 hour at room temperature. Wash the coated plate 5X with PBS-Tween/Azide and tap dry. Prepare a solution of goat anti-rabbit IgG-alkaline phosphatase that is diluted 1/5000 in PBS-Tween/Azide.
The sample containing paraquat competes with the paraquat mimic on the coating antigen for a fixed amount of the antiparaquat antibody. Quantitation of paraquat is determined by detecting the amount of antiparaquat antibody bound to the coating antigen. The bound antibody is detected using an anti-rabbit IgG antibody labeled with alkaline phosphatase (termed second antibody). The analyst is referred to Voller et al. (1976) for more details on this format. This assay has been optimized for detection of paraquat.
Environmental Immunochemical Analysis Detection of Pesticides and Other Chemicals. A User's Guide by Shirley J. Gee