Equal numbers of MG63, MG63-R1, and MG63-R2 cells (approximately 2×107 cells) were lysed in 3 mL RIPA buffer containing protease inhibitor cocktail. After centrifuging at 13,000 rpm for 15 min, 0.2 mL of supernatant was used as input and the other supernatant was subjected to IP assays using Protein-G magnetic beads coupled with anti-FOXM1, anti-CtBP1, or IgG antibodies. The input and purified protein complexes subjected to western blotting to detect FOXM1 and CtBP1 protein levels. For Co-IP assay, MG63 cells were transfected with different combinations of Flag-tagged and MYC-tagged plasmids, as indicated in the figures. Equal amounts of the transfected cells (approximately 2×107 cells) were subjected to IP procedures using Flag-agarose beads (Sigma-Aldrich, #A4596) and MYC-agarose beads (Sigma-Aldrich, #A7470). The input and purified protein complexes were subjected to western blotting using anti-Flag and anti-MYC antibodies.
Myc agarose beads
Manufactured by Merck Group
MYC-agarose beads are a laboratory product designed for protein purification. They are composed of agarose beads that have been functionalized with an anti-MYC antibody, which can be used to capture and isolate MYC-tagged proteins from cell lysates or other sample types.
Automatically generated - may contain errors
Lab products found in correlation
Flag agarose bead, by Merck Group (1 mentions)
Anti myd88 4283, by Cell Signaling Technology (1 mentions)
Mouse il 12p40 elisa kit, by R&D Systems (1 mentions)
Anti nfatc3 a303 572a, by Fortis Life Sciences (1 mentions)
Mouse ifn α elisa kit, by Thermo Fisher Scientific (1 mentions)
Anti hdac1 5356, by Cell Signaling Technology (1 mentions)
Cpg dna, by Merck Group (1 mentions)
On targetplus sirna, by GE Healthcare (1 mentions)
Ha agarose beads, by Merck Group (1 mentions)
Glutathione sepharose 4b beads, by GE Healthcare (1 mentions)
Anti ha antibody, by Merck Group (1 mentions)
Anti β actin, by Abcam (1 mentions)
2 protocols using myc agarose beads
1
Co-Immunoprecipitation of FOXM1 and CtBP1
2
Immunoblot and Immunoprecipitation Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
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CpG DNA were purchased from Sigma-Aldrich. The ON-TARGETplus siRNA were from GE Healthcare. The following antibodies were used for immunoblot analysis and/or immunoprecipitation: anti-IRF7 (sc-74472), anti-IRF7 (sc-9083), anti-NFATC1 (sc-13033), anti-NFATC2 (sc-7296), anti-NFATC3 (sc-8321), anti-NFATC4 (sc-13036), and anti-NFAT5 (sc-13035) were from Santa Cruz Biotechnology, Inc.; anti-IRF7 (51–3300) was from Thermo Fisher Scientific; anti-MyD88 (4283), anti–NF-κB1 p105/p50, and anti-HDAC1 (5356) were from Cell Signaling Technology; and anti-NFATC3 (A303-572A) was from Bethyl Laboratories, Inc. HRP-conjugated anti-HA (ab1265), anti-Myc (ab62928), and anti-transferrin receptor antibody (ab84036) were from Abcam; anti–β-actin, anti-HA antibodies, anti-GST HRP conjugate antibody, HA–agarose beads, and Myc–agarose beads were purchased from Sigma-Aldrich. Glutathione Sepharose 4B beads were from GE Healthcare. Human IFN-α, TNF-α, and IL-6 ELISA kits were from Mabtech. The mouse IFN-α ELISA kit was from eBioscience. The mouse IL-12p40 ELISA kit was from R&D Systems.
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