Primary MEFs (WT, Regnase-1–deficient, and Act1 deficient) and HEK293 cells transiently expressing FLAG- or Myc-tagged proteins were used for immunoprecipitation. Cells were disrupted by ultrasonication. After removal of cell debris by centrifugation at 20,000 ×g for 5 min, the cell lysate was incubated with either anti–Regnase-1, anti-Act1 (D-11; Santa Cruz), anti-FLAG M2, or anti-Myc (Sigma) antibodies bound to rProtein A Sepharose Fast Flow, Protein G Sepharose 4 Fast Flow (both GE Healthcare), or Dynabeads Protein G (Thermo Fisher Scientific) for 1 h at 4°C. Beads were then washed twice with Tris buffer. Immunoprecipitated proteins were eluted with 3× SDS sample buffer and subjected to 10% SDS-PAGE (PAGE).
Anti myc
Manufactured by GE Healthcare
Sourced in Sweden
Anti-Myc is a laboratory equipment product designed to detect and quantify the presence of the c-Myc protein, a transcription factor that plays a critical role in cellular processes. The core function of Anti-Myc is to provide researchers and scientists with a reliable tool for the identification and measurement of c-Myc expression in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Protein g sepharose bead, by GE Healthcare (2 mentions)
Protein g sepharose 4 fast flow, by GE Healthcare (1 mentions)
Anti flag m2, by Santa Cruz Biotechnology (1 mentions)
Rprotein a sepharose fast flow, by GE Healthcare (1 mentions)
Dynabeads protein g, by Thermo Fisher Scientific (1 mentions)
Immobilon p, by Merck Group (1 mentions)
Alexa fluor 680, by Thermo Fisher Scientific (1 mentions)
Anti flag, by Merck Group (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Irdye 800, by LI COR (1 mentions)
Anti gfp, by Roche (1 mentions)
Protein g beads, by GE Healthcare (1 mentions)
Anti flag m2 affinity gel, by Merck Group (1 mentions)
4 protocols using anti myc
1
Immunoprecipitation of Regnase-1 and Act1
2
Immunoprecipitation and Immunoblotting Protocol
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Whole-cell lysates were prepared in NP-40 buffer in native conditions as previously described (Gould et al., 1991 (link)). Proteins were immunoprecipitated from protein lysates using anti-GFP (Roche, Nutley, NJ), anti-FLAG (Sigma-Aldrich), or anti-Myc (9E10), followed by Protein G Sepharose beads (GE Healthcare).
For immunoblotting, proteins were resolved by 3–8% Tris-acetate PAGE or 4–12% NuPAGE, transferred by electroblotting to a polyvinylidene difluoride membrane (Immobilon P; Millipore, Bedford, MA), and incubated with the set of primary antibodies indicated at 1 μg/ml. Primary antibodies were detected with secondary antibodies coupled to Alexa Fluor 680 (Life Technologies, Grand Island, NY) or IRDye800 (LI-COR Biosciences, Lincoln, NE) and visualized using an Odyssey Infrared Imaging System (LI-COR Biosciences).
For immunoblotting, proteins were resolved by 3–8% Tris-acetate PAGE or 4–12% NuPAGE, transferred by electroblotting to a polyvinylidene difluoride membrane (Immobilon P; Millipore, Bedford, MA), and incubated with the set of primary antibodies indicated at 1 μg/ml. Primary antibodies were detected with secondary antibodies coupled to Alexa Fluor 680 (Life Technologies, Grand Island, NY) or IRDye800 (LI-COR Biosciences, Lincoln, NE) and visualized using an Odyssey Infrared Imaging System (LI-COR Biosciences).
3
ChIP Assay for Telomere Analysis
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The ChIP assay was performed as described previously (51 (link)). Mre11 tagged with 13Myc epitope was introduced as described (52 (link)). Immunoprecipitation of cross-linked DNA was done with anti-Myc and protein G Sepharose beads (GE healthcare). The primers targeting VI-R telomere and ARO1 (a gene far from a telomere) was used as described (13 (link)) (Supplementary Table S2 ).
4
Immunoprecipitation and ATP-Binding Assay
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Anti-FLAG (mouse monoclonal M2; Sigma Aldrich), anti-Myc (mouse monoclonal 9E10; Santa Cruz Biotechnology, Texas, USA) and anti-GST (goat polyclonal; GE Healthcare) antibodies were purchased. For IP of mysterin-3 × FLAG, the cell lysate was incubated with anti-FLAG M2 affinity gel (Sigma Aldrich). For IP of mysterin-Myc, the cell lysate was incubated with anti-Myc antibody and subsequently with protein G beads (GE Healthcare AB, Uppsala, Sweden). For ATP-binding assay, purified protein was incubated with ATP agarose (high) (Innova Biosciences, UK). The beads were repeatedly washed with lysis buffer, as described above. Subsequently, they were eluted in Laemmli's sample buffer and were analyzed using SDS-PAGE following Western blotting using the above antibodies as previously described45 (link).
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