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Running Head: LABORATORY REPORT 1 Laboratory Report Name Institution Course Date
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LABORATORY REPORT 2 Stem Cell Biology and Regenerative Medicine - Laboratory Report Methods and Materials Immunohistochemistry (IHC) Experiments: Before separating the paraffin wax from the tissue, rehydration of the tissue is performed through xylene and then a buffer solution, which includes a mixture of a conjugate base and a weak acid (ethanol, phosphate-buffered saline (PBS) as well as distal water). The experiments were conducted using rat anti-mouse F4/80 antibodies together with polyclonal rabbit anti-mouse cytokeratin at 1:150 and 1:250 dilution factors, in that order. Besides that, incubation of the conjugated secondary antibodies (biotinylated goat anti-rat and goat anti- rabbit at 1:400 for anti-F4/80 and at 1:500 for anti-CKpan, respectively) occurred. In the subsequent step, streptavidin-HRP was incubated with the aim of detecting biotinylated secondary antibodies. This was achieved by dipping the tissue in numerous ethanol solutions that had different concentrations. Xylene was utilised after the 100 percent solution dip with the aim of removing alcohol. To hold the tissue in place during imaging, it was coverslip mounted. This also helped to prevent the tissue from drying out. For tissue examination, a standard light microscope was utilised.
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