Ezview red flag m2 affinity gel

Manufactured by Merck Group

EZview Red FLAG M2 affinity gel is a pre-packed agarose-based resin designed for the purification of FLAG-tagged recombinant proteins. It features a high-affinity monoclonal anti-FLAG antibody covalently coupled to agarose beads, allowing for efficient capture and recovery of FLAG-tagged proteins.

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Lab products found in correlation

Flag peptide, by Merck Group (2 mentions) Light chain specific secondary antibody, by Jackson ImmunoResearch (1 mentions)

Close spelling variants of this product

FLAG M2 (383 citations) EZview Red ANTI-FLAG M2 Affinity Gel (119 citations) FLAG Affinity Gels (4 citations) EZview Red Anti-Flag M2 Affinity Gel clone M2 (3 citations) Affinity gels (2 citations)

3 protocols using ezview red flag m2 affinity gel

Affinity Purification of Protein Complexes

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For AP-MS, cell extracts were pre-cleared with Protein-G Dynabeads
for 1 h at 4°C before primary antibody incubation. For TAP-MS, whole
cell extracts were incubated with Strep-Tactin Sepharose (IBA) at 4°C
overnight. Cell extracts were washed three times in wash buffer and
incubated in 1× Strep-tag elution buffer (IBA) for 10 min at room
temperature on gentle vortex. The eluted solution was diluted with five
times wash buffer and incubated with EZview red Flag M2 affinity gel (Sigma)
for 4 h at 4°C. Beads were washed three times and eluted in wash
buffer containing 150 μg/mL Flag peptide (Sigma) for 30 min at room
temperature on gentle vortex.

Li C.G., Mahon C., Sweeney N.M., Verschueren E., Kantamani V., Li D., Hennigs J.K., Marciano D.P., Diebold I., Abu-Halawa O., Elliott M., Sa S., Guo F., Wang L., Cao A., Guignabert C., Sollier J., Nickel N.P., Kaschwich M., Cimprich K.A, & Rabinovitch M. (2019). PPARγ Interaction with UBR5/ATMIN Promotes DNA Repairto Maintain Endothelial Homeostasis. Cell reports, 26(5), 1333-1343.e7.

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Flag-NUSAP1 Interactome Validation

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To validate AP–MS data, cell extracts from LNCaP cells transfected with Flag-NUSAP1 were collected as described above and incubated with Ezview red Flag M2 affinity gel (Sigma Aldrich) for 4 h at 4 °C. Beads were washed three times and eluted in wash buffer containing 150 mg/mL Flag peptide (Sigma Aldrich) for 30 min at RT with a gentle vortex. For immunoblot analysis, light-chain-specific secondary antibodies (Jackson ImmunoResearch Labs, West Grove, PA, USA) were used.

Chiu C.L., Li C.G., Verschueren E., Wen R.M., Zhang D., Gordon C.A., Zhao H., Giaccia A.J, & Brooks J.D. (2023). NUSAP1 Binds ILF2 to Modulate R-Loop Accumulation and DNA Damage in Prostate Cancer. International Journal of Molecular Sciences, 24(7), 6258.

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FLAG-tagged Protein Immunoprecipitation

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Mitotic extracts were quantified and normalized to the same concentration. Approximately 90% of the extract was incubated with anti-FLAG M2 beads (Sigma, EZview RED FLAG M2 Affinity Gel) for 2 h, or with anti-FLAG (Sigma) antibody overnight and then with equilibrated protein G beads (Millipore Sigma, Protein G Agarose, Fast Flow) for 2h. After several washes with lysis buffer, samples were resuspended in 4X loading buffer (200 mM Tris-HCl, 4% SDS, 40% glycerol, 4% b-mercaptoethanol, 0.012% bromophenol blue and 50 mM DTT) and resolved by SDS-PAGE. Mouse TrueBlot ULTRA (anti-Mouse Ig (Rockland) secondary antibody) was used to probe for FLAG and CDC20 primary antibodies.

Chen O.J., Castellsagué E., Moustafa-Kamal M., Nadaf J., Rivera B., Fahiminiya S., Wang Y., Gamache I., Pacifico C., Jiang L., Carrot-Zhang J., Witkowski L., Berghuis A.M., Schönberger S., Schneider D., Hillmer M., Bens S., Siebert R., Stewart C.J.R., Zhang Z., Chao W.C.H., Greenwood C.M.T., Barford D., Tischkowitz M., Majewski J., Foulkes W.D, & Teodoro J.G. (2022). Germline Missense Variants in CDC20 Result in Aberrant Mitotic Progression and Familial Cancer. Cancer research, 82(19).

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