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Highest specificity, lowest cross-reactivity
Targets: phospho-Akt (pAkt) and human b-tubulin in T293 cells stimulated 0, 2.5, 5, or 15 min with TGF-b, phosphorylated and total protein detected.
Detection: Hybond™ ECL™ on Typhoon™ 9410 (left); LI-COR Odyssey™ scanner (right), used according to manufacturer's recommended protocols with reagents from Invitrogen and Rockland Immunochemicals Inc.*
Primary antibodies: polyclonal anti-phospho-Akt (Ser 473) and monoclonal anti-b-tubulin.
Secondary antibodies: ECL Plex™ goat-a-rabbit IgG-Cy5 and ECL Plex goat-a-mouse IgG-Cy3 (left); Alexa Fluor™ 680 goat-a-mouse IgG and IRDye™ 800 goat-a-rabbit IgG (right).
Targets: human apotransferrin four-fold dilutions starting at 5 ng, bovine cardiac muscle actin two-fold dilutions starting at 150 ng.
Detection: Hybond ECL onTyphoon 9410 (left); LI-COR Odyssey scanner (right), used according to manufacturer's recommended protocols with reagents from Invitrogen and Rockland Immunochemicals Inc.*
Primary antibodies: monoclonal anti-actin and rabbit polyclonal anti-human transferrin.
Secondary antibodies: ECL Plex goat-a-rabbit IgG-Cy5 and ECL Plex goat-a-mouse IgG-Cy3 (left); Alexa Fluor 680 goat-a-mouse IgG and IRDye 800 goat-a-rabbit IgG (right). False signals in respective channels are shown (%). |
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