Anti c myc agarose

Manufactured by Thermo Fisher Scientific

The Anti-c-Myc Agarose is a laboratory product designed for the purification and detection of proteins tagged with the c-Myc epitope. It consists of agarose beads covalently coupled with an anti-c-Myc antibody, allowing for the efficient capture and isolation of c-Myc-tagged proteins from complex samples.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Desthiobiotin, by Merck Group (1 mentions) Strep tactin superflow plus, by Qiagen (1 mentions) 3xflag peptide, by Merck Group (1 mentions) Anti flag m2 affinity resin, by Merck Group (1 mentions) Anti flag m2 affinity gel, by Merck Group (1 mentions) Np 40 lysis buffer, by Beyotime (1 mentions) Pugnac, by Merck Group (1 mentions) Anti o glcnac, by Cell Signaling Technology (1 mentions) Hrp conjugated anti β actin, by Santa Cruz Biotechnology (1 mentions) Hrp conjugated anti flag, by Merck Group (1 mentions) Gfp trap agarose, by Proteintech (1 mentions) Anti p p38, by Cell Signaling Technology (1 mentions) Anti jnk1, by Santa Cruz Biotechnology (1 mentions) Anti p65, by Cell Signaling Technology (1 mentions) Anti p stat3, by Cell Signaling Technology (1 mentions) Anti hmgb1, by Abcam (1 mentions) Anti ripk3, by Novus Biologicals (1 mentions) N acetylglucosamine glcnac, by Merck Group (1 mentions) Anti ripk1, by Cell Signaling Technology (1 mentions)

3 protocols using anti c myc agarose

Strep, Myc and FLAG Pulldown Assay

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For Strep, Myc and FLAG pulldown experiments proteins were immobilized using either Strep-Tactin Superflow Plus (QIAGEN), Anti-c-Myc Agarose (Thermo Fisher) or Anti-FLAG M2 affinity resin (Sigma). Proteins were added and incubated with the resin for 15 minutes at room temperature. Beads were washed three times with 15-20 bed volumes each. Bound proteins were eluted using either 2.5 mM desthiobiotin (Sigma), 500 μg/ml Myc peptide (GenScript) or 100 μg/ml 3X FLAG peptide (Sigma) and analyzed by SDS-PAGE.

Schreiber A., Collins B.C., Davis C., Enchev R.I., Sedra A., D’Antuono R., Aebersold R, & Peter M. (2021). Multilayered regulation of autophagy by the Atg1 kinase orchestrates spatial and temporal control of autophagosome formation. Molecular Cell, 81(24), 5066-5081.e10.

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Co-immunoprecipitation for Protein Interaction

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Co-immunoprecipitation (co-IP) analyses were performed as previously described. Briefly, plasmids were transiently transfected into HEK293 cells for 48 h. Cells were lysed using NP-40 lysis buffer (Beyotime) and IP was performed using Anti-Flag M2 Affinity Gel (Sigma), or anti-c-Myc Agarose (Thermo Fisher Scientific) and mouse IgG Agarose as a negative control (Thermo Fisher Scientific). After incubation at 4 °C for 2 h, agarose was washed three times with TBST, then incubated with 100 μl 0.1 N NH₄OH for 5 min. Following centrifugation, the resulting supernatant was collected and neutralized to pH 7.0 with using acetic acid. The samples were heated in 1 x reducing loading buffer at 95 °C for 5 min and subsequent Western Blot analysis was performed to examine protein binding. Results are representative of at least three independent experiments.

Chen J., Wo D., Ma E., Yan H., Peng J., Zhu W., Fang Y, & Ren D.N. (2019). Deletion of low-density lipoprotein-related receptor 5 inhibits liver Cancer cell proliferation via destabilizing Nucleoporin 37. Cell Communication and Signaling : CCS, 17, 174.

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Exploring Inflammatory Signaling Pathways

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LPS and CpG were from Invivogen. Pam3Cys was from EMC Microcollections. Nec-1 was from Enzo. GSK-872 and zVAD were from Millipore. S3I-201, cycloheximide, PUGNAc, ¹³C₆-glucose and N-acetylglucosamine (GlcNAc) were from Sigma-Aldrich. Lipofectamine 2000 and the Griess Reagent Kit for nitrite quantification were from Thermo Fisher Scientific. Antibodies for immunoblotting included anti-OGT, anti-O-GlcNAc, anti-mouse p-RIPK1, anti-RIPK1, anti-p-IKKα/β (S176/180), anti-p-IκBα (S32), anti-IκBα, anti-p-p65 (S536), anti-p65, anti-p-ERK1/2 (T202/Y204), anti-ERK1/2, anti-p-JNK (T183/Y185), anti-p-p38 (T180/Y182), anti-p38, anti-p-STAT3, anti-STAT3, anti-Histone H3 (Cell Signaling Technology), anti-JNK1, anti-GFP, HRP-conjugated anti-β-actin (Santa Cruz Biotechnology), HRP-conjugated anti-Flag (Sigma-Aldrich), HRP-conjugated anti-Myc (Roche), anti-IKKα, anti-NOS2, anti-GAPDH (Millipore), anti-RIPK3 (Novus Biologicals), anti-mouse MLKL (Abgent), anti-human p-RIPK3, anti-human p-MLKL, anti-mouse p-MLKL, anti-HMGB1 (Abcam), anti-IL-1α (R&D Systems) and anti-mouse p-RIPK3 (Genentech). Antibody-conjugated agarose for immunoprecipitation included anti-Flag agarose (Sigma-Aldrich), anti-c-Myc agarose (Thermo Scientific), GFP-Trap agarose (Chromotek), and sWGA agarose (Vector Laboratories).

Li X., Gong W., Wang H., Li T., Attri K.S., Lewis R.E., Kalil A.C., Bhinderwala F., Powers R., Yin G., Herring L.E., Asara J.M., Lei Y.L., Yang X., Rodriguez D.A., Yang M., Green D.R., Singh P.K, & Wen H. (2019). O-GlcNAc transferase suppresses inflammation and necroptosis by targeting receptor-interacting serine/threonine-protein kinase 3. Immunity, 50(3), 576-590.e6.

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