Immunoprecipitation of extracts from COS cells transfected with Myc-Chorionic Somatomammotropin Hormone-Like 1 (CSL) protein using Myc-Tag (9B11) Mouse mAb (Biotinylated) #2084 and Immobilized Streptavidin (Bead Conjugate) (Lane 1) shows immunocomplexes pulled down using Streptavidin (Sepharose® Bead Conjugate). Lysate and beads alone are shown in lane 2 indicating the specificity of the streptavidin beads. Western blotting was performed using Myc-Tag (9B11) Mouse mAb (HRP Conjugate) #2040.
Immunoprecipitation of extracts from NIH/3T3 cells using PTEN (138G6) Rabbit mAb (Biotinylated) #9583 and Streptavidin (Sepharose® Bead Conjugate) (Lane 1). Lysate and beads alone are shown in lane 2 indicating the specificity of the streptavidin beads. Western blotting was performed using PTEN (26H9) Mouse mAb #9556.
Immunoprecipitation of extracts from Jurkat cells using Phospho-Akt (Thr308) (244F9) Rabbit mAb (Biotinylated) #3454 and Streptavidin (Sepharose® Bead Conjugate) (Lane 1). Lysate and beads alone are shown in lane 2 indicating the specificity of the streptavidin beads. Western blotting was performed using Phospho-Akt (Thr308) (L32A4) Mouse mAb #5106.
Add 10 μl of well-vortexed beads to 200 μl of cell lysate at 1 mg/ml pre-incubated with biotinylated primary antibody. See protocol for more details.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol. Store at –20°C. Do not aliquot the antibodies.
Streptavidin (Sepharose® Bead Conjugate) is useful for the precipitation of biotinylated proteins (1,2). Recombinant streptavidin is immobilized via covalent binding of primary amino groups to N-hydroxysuccinimide (NHS)-activated Sepharose® beads.
Streptavidin is a 53,000 dalton tetrameric protein purified from the bacterium Streptomyces avidinii (3). Each subunit binds to biotin with extremely high affinity. Because of its strong non-covalent interaction with biotin, streptavidin can be used to isolate biotinylated proteins (1,2).
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