HEK-293T cells were transfected with IFI16-Myc and MAVS-3×Flag, MAVS truncations plasmids or control vector. At 48 hpt, the cells were lysed in IP-lysis buffer (Sigma-Aldrich) containing protease inhibitor cocktail (Roche) for 1 h at 4 °C. The lysates were centrifuged at 12,000× g for 15 min at 4 °C, and the supernatants were pre-cleared with mouse IgG-Agarose (Sigma-Aldrich) at 4 °C for 2 h. Then, the pre-cleared supernatants were incubated with anti-c-Myc affinity gel beads or anti-Flag affinity gel beads (Sigma-Aldrich) for 4 h or overnight at 4 °C. The precipitates were washed five times with TBS buffer and detected by western blotting.
Anti c myc affinity gel beads
Manufactured by Merck Group
Anti-c-Myc affinity gel beads are a laboratory product used to purify and isolate c-Myc tagged proteins from cell lysates or other biological samples. The beads are coated with an antibody that specifically binds to the c-Myc epitope tag, allowing for the capture and recovery of c-Myc tagged proteins.
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Lab products found in correlation
Mouse igg agarose, by Merck Group (2 mentions)
Anti flag affinity gel beads, by Merck Group (2 mentions)
Ip lysis buffer, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Recombinant human ifn β, by Thermo Fisher Scientific (1 mentions)
Mouse monoclonal anti c myc antibody, by Merck Group (1 mentions)
Mouse monoclonal anti flag m2 antibody, by Merck Group (1 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Polyinosinic polycytidylic acid poly 1 c, by Merck Group (1 mentions)
Prrsv n protein antibody, by GeneTex (1 mentions)
Mouse anti β actin monoclonal antibody, by GenScript (1 mentions)
Secondary antibodies conjugated to hrp, by Jackson ImmunoResearch (1 mentions)
2 protocols using anti c myc affinity gel beads
1
Immunoprecipitation of IFI16 and MAVS
2
Comprehensive Protein Immunoprecipitation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Mouse anti-Flag M2 monoclonal antibody, mouse anti-c-Myc monoclonal antibody, anti-Flag affinity gel beads, anti-c-Myc affinity gel beads, and mouse IgG agarose were purchased from Sigma-Aldrich. PRRSV N protein antibody was purchased from GeneTex. Mouse anti-β-actin monoclonal antibody was purchased from GenScript. Rabbit anti-MAVS monoclonal antibody was purchased from Cell Signaling Technology (CST). The secondary antibodies conjugated to HRP were purchased from Jackson Immuno Research. Polyinosinic-polycytidylic acid (polyI:C) was purchased from Sigma-Aldrich. Recombinant human IFN-β was purchased from PEPROTECH. Lipofectamine2000 transfection reagent and Lipofectamine RNAiMAX transfection reagent were purchased from Invitrogen (Carlsbad, CA, USA).
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