Gfp trap

Manufactured by Merck Group

GFP-trap is a laboratory equipment product for the purification and isolation of Green Fluorescent Protein (GFP) and GFP-fusion proteins. It utilizes a single-domain antibody fragment coupled to agarose beads to specifically capture and immobilize GFP-tagged proteins from complex samples.

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Lab products found in correlation

Gfp antibody, by Merck Group (1 mentions) Anti flag m2 resin, by Merck Group (1 mentions) Nupage, by Thermo Fisher Scientific (1 mentions) Protease and phosphatase inhibitors cocktail, by Roche (1 mentions)

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GFP-Trap beads (5 citations)

2 protocols using gfp trap

Verifying GFPMBP-BRCA2 Protein Expression

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To verify protein expression, a fraction of the cells was harvested and lysed in lysis buffer containing 50 mM Hepes (pH 7.5), 250 mM NaCl, 5 mM EDTA, 1% NP-40, 1 mM dithiothreitol (DTT), 1 mM PMSF, and Protease Inhibitor Cocktail (Roche). GFPMBP-BRCA2 protein was detected from GFP-trap (ChromoTek) immunoprecipitates by immunoblotting with GFP antibody (Sigma G1544, 1:500).

Caputo S.M., Léone M., Damiola F., Ehlen A., Carreira A., Gaidrat P., Martins A., Brandão R.D., Peixoto A., Vega A., Houdayer C., Delnatte C., Bronner M., Muller D., Castera L., Guillaud-Bataille M., Søkilde I., Uhrhammer N., Demontety S., Tubeuf H., Castelain G., Jensen U.B., Petitalot A., Krieger S., Lefol C., Moncoutier V., Boutry-Kryza N., Nielsen H.R., Sinilnikova O., Stoppa-Lyonnet D., Spurdle A.B., Teixeira M.R., Coulet F., Thomassen M, & Rouleau E. (2018). Full in-frame exon 3 skipping of BRCA2 confers high risk of breast and/or ovarian cancer. Oncotarget, 9(25), 17334-17348.

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Immunoprecipitation of Protein Complexes

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Whole cells protein extracts were generated using a lysis buffer containing 150 mM NaCl, 20 mM Tris-HCl (pH 7.65), 0.5% NP-40, 5 mM EDTA, 5% glycerol and protease and phosphatase cocktail inhibitors (Roche). 30 μg of lysates measured by Bradford assay were loaded into precast gels (Nu-PAGE Invitrogen, 4-12%). Immunoprecipitation was carried out by incubating 1 mg of protein lysate with Anti-Flag M2 resin (sigma) or GFP-trap, overnight at 4 °C. The following day, the beads were washed [20 mM Tris-HCl (pH 7.65), 150 mM NaCl, 3 mM MgCl_2, 10% Glycerol, 0.01% NP-40] and eluted in either glycine or 4x SDS loading buffer. With GFP-trap, proteins were eluted directly in 4x loading buffer.

Parnandi N., Rendo V., Cui G., Botuyan M.V., Remisova M., Nguyen H., Drané P., Beroukhim R., Altmeyer M., Mer G, & Chowdhury D. (2021). TIRR inhibits the 53BP1-p53 complex to alter cell-fate programs. Molecular cell, 81(12), 2583-2595.e6.

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