|Transfected Only||Mouse IgG2a|
Immunoprecipitation of extracts from COS-7 cells, mock transfected (-) or transfected with a construct overexpressing GST-TrCP1 protein (GST-TrCP1; +), using GST (26H1) Mouse mAb (Magnetic Bead Conjugate) (lanes 5 and 6) and Mouse IgG (Magnetic Bead Conjugate) #5873 (lanes 3 and 4). Lanes 1 and 2 show 10% lysate input. Western blot analysis was performed using GST (91G1) Rabbit mAb #2625.Learn more about how we get our images.
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
Supplied in PBS Buffer (pH 7.2), 0.1% Tween® 20. Store at 4°C. Do not aliquot the antibodies.
GST (26H1) Mouse mAb (Magnetic Bead Conjugate) detects transfected glutathione S-transferase (GST) fusion proteins.
All Species Expected
Monoclonal antibody is produced by immunizing animals with a GST fusion protein.
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.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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