Immunoprecipitation of COS-7 cell extracts, untransfected or transfected with Myc-Chorionic Somatomammotropin Hormone-Like 1 (Myc-CSL) protein, using Myc-Tag (9B11) Mouse mAb (Magnetic Bead Conjugate). Blots were probed with Myc-Tag (71D10) Rabbit mAb #2278.
This Cell Signaling Technology antibody is immobilized by the covalent reaction of hydrazinonicotinamide-modifed antibody with formylbenzamide-modified magnetic bead. Myc-Tag (9B11) Mouse mAb (Magnetic Bead Conjugate) is useful for immunoprecipitation assays of Myc epitope-tagged proteins.
Supplied in PBS Buffer (pH 7.2), 0.1% Tween® 20. Store at 4°C. Do not aliquot the antibodies.
This protocol is intended for immunoprecipitation of native proteins for analysis by western immunoblot or kinase activity.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
10X Cell Lysis Buffer: (#9803) 20 mM Tris (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100, 2.5 mM Sodium pyrophosphate, 1 mM β-glycerophosphate, 1 mM Na3VO4, 1 μg/ml Leupeptin
NOTE: CST recommends adding 1 mM PMSF (#8553) before use*.
Proceed to one of the following specific set of steps.
NOTE: To minimize masking caused by denatured IgG heavy chains (~50 kDa), we recommend using Mouse Anti-Rabbit IgG (Light-Chain Specific) (L57A3) mAb (#3677) or Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127). To minimize masking caused by denatured IgG light chains (~25 kDa), we recommend using Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (#3678) (or HRP conjugate #5127).
posted December 2007
Protocol Id: 408
Myc-Tag (9B11) Mouse mAb (Magnetic Bead Conjugate) detects recombinant proteins containing the Myc epitope tag. The antibody recognizes the Myc-tag fused to either the amino or carboxy terminus of targeted proteins in transfected cells.Species Reactivity:
All Species Expected
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues 410-419 of human c-Myc (EQKLISEEDL).
Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation, and immunostaining techniques. Because of their small size, they are unlikely to affect the tagged protein’s biochemical properties.
The Myc epitope tag is widely used to detect expression of recombinant proteins in bacteria, yeast, insect, and mammalian cell systems (1).
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