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 Post subject: The slides were then incubated with primary antibody overnig
PostPosted: Mon Jan 22, 2018 3:18 am 
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Joined: Wed Jul 31, 2013 1:53 am
Posts: 205
The next day the slides were incubated with biotinylated secondary antibody, washed again with PBS, then incubated with streptavidin conjugated to horseradish-peroxidase. After a final PBS wash, a drop of freshly prepared DAB was applied on the slide for color development. The reaction was stopped when a uniform brown color became visible on the slide by rinsing in running water. Counterstaining was done with hematoxylin for 5–10 seconds. A control experiment was performed by replacing primary antibody Tubacin|Tubulin Acetylation Inducer HDAC inhibitor with PBS. This method was a modification at the manufacturers’ instructions. The evaluation of Von Willbrand Factor expression and marrow microvessel density were determined by counting the number of endothelial cells showing positive staining Von Willbrand Factor, and the MVD was determined by the total number of vWF TWS119 GSK-3 inhibitor stained cells divided by the measured area in previous studies.


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