Streptavidin irdye 800

Manufactured by Rockland Immunochemicals

Streptavidin-IRDye 800 is a fluorescent conjugate used for detection and quantification in various bioanalytical techniques. Streptavidin, a protein derived from the bacterium Streptomyces, is covalently linked to the near-infrared dye IRDye 800. This conjugate can be used to label and detect biotinylated biomolecules, such as proteins, nucleic acids, and other targets.

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Lab products found in correlation

Streptavidin agarose, by Thermo Fisher Scientific (1 mentions) Mouse anti hsp70, by Abcam (1 mentions) Mouse anti actin, by Merck Group (1 mentions) Mouse anti tubulin, by Abcam (1 mentions) Anti bip, by Abcam (1 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Anti flag, by Merck Group (1 mentions)

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IRDye 800 (95 citations)

2 protocols using streptavidin irdye 800

AHA-Labeling and Biotin Enrichment of Newly Synthesized Proteins

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To assess the effect of 4E-BP overexpression, 4E-BP^LLAA was induced in S2 cells stably transfected with pMT-4E-BP^LLAA with 0.5mM copper sulfate for 2 h in methionine-free Schneider’s medium to enable AHA uptake. The cells were then labeled with 4 mM AHA (Anaspec) for 1 h. AHA-labeled proteins in the extracts were clicked to biotin-PEG4 alkyne (Invitrogen) using the manufacturer’s protocols. AHA-conjugated peptides were further enriched by incubation with Streptavidin-agarose (Thermo Fisher Scientific) followed by rigorous washing with PBS-Tween (0.5%). The bound fraction was collected by boiling the beads in sample buffer and analyzed by Western blotting with Streptavidin-IRDye 800 (Rockland Immunochemicals), guinea pig anti-BiP (1:1,000), mouse anti-Hsp70 (Abcam), mouse antiactin (EMD Millipore), and mouse antitubulin (Abcam).

Kang M.J., Vasudevan D., Kang K., Kim K., Park J.E., Zhang N., Zeng X., Neubert T.A., Marr MT I.I, & Ryoo H.D. (2017). 4E-BP is a target of the GCN2–ATF4 pathway during Drosophila development and aging. The Journal of Cell Biology, 216(1), 115-129.

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Isolation and Analysis of Ribosome-Bound Proteins

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HEK293T cells transiently transfected with N-3XFlag-ADAMTS13 plasmid and labeled with 200 μM 6AF for 24 h, then treated with 100 μM cycloheximide (Sigma) for 15 min at 37°C. The cells were then lysed in 50 mM NH₄HCl, 12 mM MgCl₂, 50 mM Tris acetate and 0.5% NP-40. The lysates were passed through a 27.5 gauge syringe twice to thoroughly free any adherent ribosomes and debris is removed by centrifuging at 12,000 g (20 min at 4°C). The lysate was “clicked” (as described above) and loaded on a 34% sucrose cushion containing 50 mM NH₄HCl, 12 mM MgCl₂, 50 mM Tris acetate and centrifuged at 100,000 g for 3 h. The pellet was rinsed 3X with lysis buffer, resuspended in sample buffer, and analysed by western blotting (anti-Flag (Sigma), Streptavidin IRDye 800 (Rockland Immunochemicals)), and imaging performed with the Odyssey system.

Vasudevan D., Takeuchi H., Johar S.S., Majerus E, & Haltiwanger R.S. (2014). Peters plus syndrome mutations disrupt a non-canonical ER quality control mechanism. Current biology : CB, 25(3), 286-295.

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