Giardia cells carrying pGlPLK-HA.neo and pGlKin-13-Myc.pac were harvested, washed 3 times with ice-cold PBS, and lysed with lysis buffer (10 mM Tris-Cl, 5 mM EDTA, 130 mM NaCl, and 1% Triton X-100, pH 7.4) containing protease inhibitor cocktail (GenDEPOT, Katy, Taxas, USA). Lysates precleared using Protein A/G Agarose (Thermo Fisher Scientific) for 1 h at 4°C were incubated with monoclonal mouse anti-HA agarose beads (Sigma-Aldrich) or monoclonal mouse anti-Myc antibodies (Clontech, Mountain View, California, USA) at 4°C overnight. Beads were washed with immunoprecipitation washing buffer (50 mM Tris-Cl, 150 mM NaCl, and 1% Triton X-100, pH 7.4) and resuspended in 2×SDS sample buffer. The samples were analyzed by Western blotting using anti-HA or anti-Myc antibodies.
Monoclonal mouse anti ha agarose beads
Manufactured by Merck Group
Sourced in United States
Monoclonal mouse anti-HA agarose beads are a laboratory reagent used for the purification and detection of proteins tagged with the HA (Hemagglutinin) epitope. The beads are coated with a monoclonal antibody specific to the HA tag, allowing for the efficient capture and isolation of HA-tagged proteins from complex mixtures.
Automatically generated - may contain errors
2 protocols using monoclonal mouse anti ha agarose beads
1
Immunoprecipitation of Giardia Proteins
2
Giardia CDK and Cyclin Complexes
Check if the same lab product or an alternative is used in the 5 most similar protocols
Giardia cells carrying one of the two CDK plasmids (pGlCDK1HA.NEO and pGlCDK2HA.NEO) and one of the nine cyclin plasmids were harvested, washed twice with ice-cold PBS, and lysed in protein lysis buffer (10 mM Tris-HCl, 5 mM EDTA, 130 mM NaCl, and 1% Triton X-100, pH 7.4) containing a protease inhibitor cocktail (GenDEPOT). Lysates were precleared with protein A/G agarose (Thermo Fisher Scientific) for 1 h at 4°C. Subsequently, precleared lysates were incubated with monoclonal mouse anti-HA agarose beads (Sigma-Aldrich) or monoclonal mouse anti-Myc antibodies (Clontech) at 4°C overnight. The beads were washed twice with co-IP washing buffer (50 mM Tris-HCl, 150 mM NaCl, and 1% Triton X-100, pH 7.4) for 10 min and resuspended in 2 × SDS sample buffer. The precipitates were then analyzed by Western blotting using anti-HA or anti-Myc antibodies.
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