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Eva Wahlund
Matilda Ahnfelt (technician), Dijana Cerjan (technician), Åsa Edvinsson (technician), Inga Hallin (technician) and Urban Ryberg (technician).
(i) Handling and storage of antisera, (ii) testing and titration of affinity purified monospecific antibodies and collaborative antibodies, (iii) immunostainings on full-scale TMAs for all approved and validated antibodies, (iv) protein extractions for western blot analysis, (v) western blot analysis of external antibodies, (vi) culturing of cells used for cell TMAs and lysates for Western blot analysis.
Antibodies are titrated using a specially designed test-TMA, representing a limited selection of tissues and cells. The titration is done according to a defined schedule where different antibody dilutions and antigen retrieval methods are tested. The primary working dilution is based on the protein concentration for each antibody. Automated immunostaining instruments and commercially available detection kits are used to ensure standardization and reproducibility.
In addition protein extraction is done from cultured cell lines and fresh frozen tissues and used for Western blots.
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