title: Clean Western blot signals from immunoprecipitated samples.
abstract: We present a strategy that overcomes the high background arising during Western blotting (WB) detection of proteins obtained through immunoprecipitation (IP). Traditional HRP-conjugated secondary antibodies, which detect the denatured heavy and light antibody chains, produce high background that often mask the signals of interest on WBs. Here, we show that HRP-conjugated Protein A and Protein G, which detect almost exclusively intact antibody molecules, can be effectively used to obtain clean and specific WB signals of target proteins.
authors.affiliation: Laboratory of Cellular and Molecular Biology, National Institute on Aging-IRP, National Institutes of Health, Box 12, 5600 Nathan Shock Dr., Baltimore, MD 21224, USA. ashishl@grc.nia.nih.gov
authors.forename: Ashish
authors.initials: A
authors.lastname: Lal
authors.forename: Susan R
authors.initials: SR
authors.lastname: Haynes
authors.forename: Myriam
authors.initials: M
authors.lastname: Gorospe
mesh_terms: D015153:Blotting, Western / Q000379:methods*; D006735:Horseradish Peroxidase / Q000737:chemistry; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000378:metabolism; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000378:metabolism; D047468:Immunoprecipitation / Q000379:methods*; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000378:metabolism
chemical_list: C029152:G-substrate; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D009419:Nerve Tissue Proteins; D013205:Staphylococcal Protein A; D006735:Horseradish Peroxidase