Immunoprecipitation is a widely used technique to enrich proteins, peptides, and protein complexes. This video includes a step-by-step demonstration of the Immunoprecipitation protocol for Native Protein, performed by scientists at Cell Signaling Technology.
● CST Immunoprecipitation Protocols: [ Ссылка ]
● Immunoprecipitation Resource Center: [ Ссылка ]
● Immunoprecipitation Experimental Design Tips webpage: [ Ссылка ]
● CST Proteomics Analytics Services: [ Ссылка ]
Contents:
0:00 Introduction
2:13 Protocol Considerations
3:30 Antibody Selection for Single-Protein IP
4:30 Antibody Selection for Western Blot analysis of IP
4:53 Choice of Beads
6:08 Controls for IP experiments
6:29 Input Control
6:49 Isotype Control
7:12 Bead-Only Control and Unbound Fraction Control
7:46 Lysate Considerations
8:42 A. Solutions and Reagents
10:21 B. Preparing Cell Lysates
12:20 C. Immunoprecipitation
13.37 Lysate Pre-Clearing
17:10 IP Reaction Options
17:32 Unconjugated or Biotinylated Antibodies
18:26 Biotinylated Antibodies + Streptavidin Sepharose Beads
18:44 Unconjugated Antibodies + Magnetic Beads
19:14 Immobilized Antibody-Bead Conjugates
19:46 Tips for Washing Enriched immunocomplexes
22:22 D. Sample Analysis
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