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Immobilized Streptavidin (Bead Conjugate) #3419
Protocols
- 3419:
- Immunoprecipitation
Description
Immobilized Streptavidin (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.
IP
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 Immobilized Streptavidin (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.
IP
Immunoprecipitation of extracts from NIH/3T3 cells using PTEN (138G6) Rabbit mAb (Biotinylated) #9583 and Immobilized Streptavidin (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.
IP
Immunoprecipitation of extracts from Jurkat cells using Phospho-Akt (Thr308) (244F9) Rabbit mAb (Biotinylated) #3454 and Immobilized Streptavidin (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.
Directions for Use
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.
Background
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).
- Updyke, T.V. and Nicolson, G.L. (1984) J Immunol Methods 73, 83-95.
- Buckie, J.W. and Cook, G.M. (1986) Anal Biochem 156, 463-72.
- Chaiet, L. and Wolf, F.J. (1964) Arch Biochem Biophys 106, 1-5.
- Reznik, G.O. et al. (1998) Proc Natl Acad Sci USA 95, 13525-13530.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
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- 9583 PTEN (138G6) Rabbit mAb (Biotinylated)
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 7072 Phototope®-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
