By Simon M.I., Thorner J., Emr S.D.
The seriously acclaimed laboratory normal for greater than 40 years, equipment in Enzymology is among the such a lot hugely revered guides within the box of biochemistry. due to the fact 1955, each one quantity has been eagerly awaited, usually consulted, and praised via researchers and reviewers alike. Now with greater than three hundred volumes (all of them nonetheless in print), the sequence includes a lot fabric nonetheless proper this present day - actually a necessary booklet for researchers in all fields of existence sciences.
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Additional resources for Applications of Chimeric Genes and Hybrid Proteins: Cell Biology and Physiology
0, to stop the reaction. Incubate on a rotator at 4” for 4 hr. e. 0)]. f. Store the beads in modified RIPA buffer in a tightly closed tube at 4”. El AP FUSIONPRO-~N~A~ in Situ PROBES 29 2. For a six-well tissue culture plate, label the cells with 2 ml of labeling solution (DMEM without methionine, containing 10% [v/v] dialyzed serum, and 400 &i of [35S]methionine) at 37” for 3 to 6 hr. 3. don a Centricon-10 (Amicon). 4. Mix the supernatant with 20 ~1 of beads coupled with anti-AP antibody for 30 min or a rotator at room temperature.
For library screening it is important to ensure a uniform density of cells over all parts of the plate, and to stain cells that are just under confluence, or just recently confluent. Overconfluent cells can pile up, trapping the fusion protein probe and sometimes causing unpredictable background staining. Also note that during washing procedures, cells can dry and fall off quickly if all the medium is siphoned out of the plate. The problem is mainly seen around the edges, and so is more severe in smaller plates.
Chem. 265, lb996 i1990). HRP 131 CHIMERAS AS EM TRACERS 39 The HRP and c-myc KDEL fragments are ligated together and the product separated by electrophoresis on low gelling temperature (LGT) agarose. This product is then ligated into PSR WSS. Sequences of the human transferrin receptor, which include only the transmembrane domain and the cytoplasmic domain or only the transmembrane domain plus residues on either side of the signal anchor (residues 60-91) are generated by PCR from cDNA provided by I.
Applications of Chimeric Genes and Hybrid Proteins: Cell Biology and Physiology by Simon M.I., Thorner J., Emr S.D.