Flag beads

Manufactured by Thermo Fisher Scientific

Flag-beads are uniform, paramagnetic beads coated with a specific antibody or protein. They are designed for the capture, detection, and enrichment of target molecules in various analytical and research applications.

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Lab products found in correlation

Nanoacquity hplc pump, by Waters Corporation (1 mentions) Ltq orbitrap elite, by Thermo Fisher Scientific (1 mentions) Anti flag, by Merck Group (1 mentions) Benzonase, by Merck Group (1 mentions) Anti gfp, by Roche (1 mentions)

Close spelling variants of this product

Anti-Flag magnetic beads Anti-FLAG M2 agarose beads Anti-FLAG M2 beads Flag magnetic beads Anti-FLAG L5 Magnetic Beads Anti-FLAG beads Flag M2 beads Anti-FLAG M2 Magnetic Beads

2 protocols using flag beads

Proteomic Profiling of Flag-USP21 Interactome

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The Flag-USP21 were transfected into T24 cells. Anti-Flag affinity magnetic beads were used for immunoprecipitation. The proteins were then identified and subjected to mass spectrometry as previously described [26 ,27 ]. Briefly, protein extracted from H1299 cells was used as input control and for immunoprecipitation. Sample was incubated with anti-Flag antibody and 50 μl agarose A protein overnight at 4 °C. The Flag-beads (Thermo Fisher Scientific) were washed with lysis buffer and boiled in SDS sample buffer, and the pulled-down protein was separated on SDS-PAGE gels. Then, the gels were stained with Coomassie Blue, and differentially abundant bands were cut out for mass spectrometry using LTQ Orbitrap Elite (Thermo Fisher) and a Waters NanoAcquity HPLC pump (Milford, MA, USA).

Ma Q., Wu F., Liu X., Zhao C., Sun Y., Li Y., Zhang W., Ju H, & Wang Y. (2024). 20-hydroxyecdysone suppresses bladder cancer progression via inhibiting USP21: A mechanism associated with deubiquitination and degradation of p65. Translational Oncology, 45, 101958.

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Investigating SDC1-Flag Interactions

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HeLa cells cultured in 6-well plates were transfected with a SDC1-Flag construct (WT, TMD_L, or HS) and GFP-LPL or with GFP-LPL alone. After 24h, cells were washed with 1x dPBS and harvested by scraping in 1mL dPBS. Cells were collected by centrifugation at 1,500 × g and lysed in buffer containing 50mM Tris pH 8.0, 150mM NaCl, 0.5% Triton X-100, and 250 U/mL benzonase (Sigma, E1014). Cell lysate was clarified by centrifugation at 14,000 × g. Protein concentrations were measured by Bradford and volumes were adjusted for equal protein concentrations. 200 μl cell lysate was added to 10 μl Flag beads (ThermoFisher, 36797) and incubated for 1h at 37 °C. Beads were washed 3x in lysis buffer and protein was eluted by boiling in SDS page protein loading buffer. 5% input and 50% IP are loaded in gel for Western Blot. Proteins were detected with anti-Flag (1:5000; Sigma, 1804) and anti-GFP (1:1000; Roche, 11814460001) antibodies.

Sundberg E.L., Deng Y, & Burd C.G. (2019). Syndecan-1 mediates sorting of soluble lipoprotein lipase with sphingomyelin-rich membrane in the Golgi apparatus. Developmental cell, 51(3), 387-398.e4.

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