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DyLightTM dyes can be used for two color Western Blot detection with low background and high signal. Anti tubulin was detected using a DyLightTM 549 conjugate. Anti TNFa was detected using a DyLightTM 649 conjugate. The image was captured using the TyphoonTM 9410 Imaging System.
Properties of DyLightTM Fluorescent Dyes.
Western Blot of Rabbit Anti-HA Epitope Tag Antibody Dylight?649 Conjugated. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: Flag Lysate (p/n W00-E01-383). Lane 3: 12 Epitope GST Tag Protein Marker Lysate (p/n MB-302-0100). Lane 4: HeLa Whole Cell Lysate (p/n W09-000-364). Load: 10μg/lane. Primary Antibody: Anti-HA Epitope Tag Dylight?649 at 1.0μg/mL overnight at 2-8°C. Blocking: MB-070 Buffer at RT for 1hr. Expected MW: ~48kDa for Flag Lysate, ~48kDa for 12 Epitope GST Tag. Exposure: 0.33 sec.
Western Blot of DylightTM 649 conjugated Rabbit anti-HA antibody. Lane 1: Epitope Tag Protein Marker Lysate (p/n MB-301-0100). Primary antibody: none. Secondary antibody: DylightTM 649 rabbit secondary antibody at 1:1,000 for 60 min at RT. Block: MB-070 for 30 min at RT. Predicted/Observed size: ~68 kDa/~68 kDa for HA Epitope Tag.
DyLight? dyes can be used for two-color Western Blot detection with low background and high signal. Anti-tubulin was detected using a DyLight? 549 conjugate. Anti-TNFa was detected using a DyLight? 649 conjugate. The image was captured using the Typhoon? 9410 Imaging System.
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