Cells were harvested with lysis buffer (150 mM NaCl, 1% NP-40, 2 mM EDTA, 50 mM Tris, pH 8.0, and protease inhibitor cocktail). Approximately 1 mg of whole cell lysate was incubated with 1 μg of the indicated Ab with constant agitation overnight at 4°C. Then, 30 μL protein G agarose bead slurry (Sigma-Aldrich, P3296) was added to pull down the immunocomplexes. The beads were collected by centrifugation, washed extensively with lysis buffer 3–5 times, and boiled with 2× SDS loading buffer. The protein samples were then subjected to immunoblot analysis with the appropriate Ab.
Protein g agarose bead slurry
Manufactured by Merck Group
Protein G agarose bead slurry is a laboratory product that consists of Protein G, a bacterial-derived immunoglobulin-binding protein, immobilized on agarose beads in an aqueous suspension. The core function of this product is to provide a matrix for the capture and purification of antibodies from biological samples.
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Lab products found in correlation
2 protocols using protein g agarose bead slurry
1
Immunoprecipitation Protein Extraction
2
Immunoprecipitation and Immunoblot Analysis
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Cells were harvested with lysis buffer (150 mM NaCl, 1% NP-40, 2 mM EDTA, 50 mM Tris, pH 8.0, and protease inhibitor cocktail). Approximately 1 mg of whole cell lysate was incubated with 1 μg of the indicated antibody with constant agitation overnight at 4°C. 30 μl protein G agarose bead slurry (Sigma; P3296) was added to pull down the immunocomplexes. The beads were collected by centrifugation, washed extensively with lysis buffer 3–5 times, and boiled with 2× SDS loading buffer. The protein samples were then subjected to immunoblot analysis with the appropriate antibody.
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