ARPE-19 cells (5 million) were metabolically labeled with 35S-methionine for 6 h at 37 °C. Cells were lysed in 0.5% NP40 lysis mix26 (link) and immunoprecipitated using mAbs 2E7 and 12H8. Following incubation of immune complexes with protein G-agarose beads (RepliGen), the recovered polypeptides were resolved on a SDS-polyacrylamide gel (12.5%) and visualized using fluorography26 (link).
Protein g agarose beads
Manufactured by Repligen
Protein G-agarose beads are a solid-phase affinity chromatography material used for the purification of immunoglobulins and other proteins that bind to Protein G. Protein G is a bacterial cell wall protein that has a high affinity for the Fc region of most immunoglobulin classes and subclasses from various species.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using protein g agarose beads
1
Immunodetection of Labeled Proteins in ARPE-19 Cells
2
Identification of 2E7 mAb Target Protein
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For identifying the target protein of mAb 2E7, ARPE-19 cells (10 million) lysed with 0.5% NP40 lysis mix26 (link) were incubated with 2E7 mAb pre-bound to protein G-agarose beads (RepliGen) for 6 h at 4 °C. The recovered proteins were resolved on a SDS-polyacrylamide gel (12.5%) and visualized using Gel Code Blue Reagent (Thermo Fisher). Polypeptides migrating at 60 kDa and 50 kDa were excised and subjected to mass spectrometry proteomic analysis at Bioproximity, Inc (Chantilly, VA)31 (link).
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